<?xml version="1.0" encoding="US-ASCII" standalone="no"?>
    <!-- Field: Doc-Info; Name: Generator; Value: GoFiler Complete; Version: 6.5a -->
    <!-- Field: Doc-Info; Name: VendorURI; Value: https://www.novaworks.com -->
    <!-- Field: Doc-Info; Name: Status; Value: 0x00000000 -->
<link:linkbase xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:link="http://www.xbrl.org/2003/linkbase" xmlns:xbrldt="http://xbrl.org/2005/xbrldt" xmlns:xbrli="http://www.xbrl.org/2003/instance" xsi:schemaLocation="http://www.xbrl.org/2003/linkbase http://www.xbrl.org/2003/xbrl-linkbase-2003-12-31.xsd">
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#Cover" roleURI="http://genelux.com/role/Cover" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BalanceSheets" roleURI="http://genelux.com/role/BalanceSheets" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BalanceSheetsParenthetical" roleURI="http://genelux.com/role/BalanceSheetsParenthetical" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StatementsOfOperationsAndComprehensiveLoss" roleURI="http://genelux.com/role/StatementsOfOperationsAndComprehensiveLoss" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StatementsOfStockholdersEquity" roleURI="http://genelux.com/role/StatementsOfStockholdersEquity" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StatementsOfCashFlows" roleURI="http://genelux.com/role/StatementsOfCashFlows" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BasisOfPresentation" roleURI="http://genelux.com/role/BasisOfPresentation" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SummaryOfSignificantAccountingPolicies" roleURI="http://genelux.com/role/SummaryOfSignificantAccountingPolicies" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#LicenseAgreements" roleURI="http://genelux.com/role/LicenseAgreements" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#FairValueMeasurements" roleURI="http://genelux.com/role/FairValueMeasurements" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#Investments" roleURI="http://genelux.com/role/Investments" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BalanceSheetAccounts" roleURI="http://genelux.com/role/BalanceSheetAccounts" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#Leases" roleURI="http://genelux.com/role/Leases" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#CommitmentsAndContingencies" roleURI="http://genelux.com/role/CommitmentsAndContingencies" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StockholdersEquity" roleURI="http://genelux.com/role/StockholdersEquity" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StockBasedCompensation" roleURI="http://genelux.com/role/StockBasedCompensation" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#NetLossPerShare" roleURI="http://genelux.com/role/NetLossPerShare" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SegmentInformation" roleURI="http://genelux.com/role/SegmentInformation" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SubsequentEvents" roleURI="http://genelux.com/role/SubsequentEvents" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SummaryOfSignificantAccountingPoliciesPolicies" roleURI="http://genelux.com/role/SummaryOfSignificantAccountingPoliciesPolicies" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#FairValueMeasurementsTables" roleURI="http://genelux.com/role/FairValueMeasurementsTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#InvestmentsTables" roleURI="http://genelux.com/role/InvestmentsTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BalanceSheetAccountsTables" roleURI="http://genelux.com/role/BalanceSheetAccountsTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#LeasesTables" roleURI="http://genelux.com/role/LeasesTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StockholdersEquityTables" roleURI="http://genelux.com/role/StockholdersEquityTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StockBasedCompensationTables" roleURI="http://genelux.com/role/StockBasedCompensationTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#NetLossPerShareTables" roleURI="http://genelux.com/role/NetLossPerShareTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SegmentInformationTables" roleURI="http://genelux.com/role/SegmentInformationTables" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BasisOfPresentationDetailsNarrative" roleURI="http://genelux.com/role/BasisOfPresentationDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SummaryOfSignificantAccountingPoliciesDetailsNarrative" roleURI="http://genelux.com/role/SummaryOfSignificantAccountingPoliciesDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#LicenseAgreementsDetailsNarrative" roleURI="http://genelux.com/role/LicenseAgreementsDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfFinancialAssetsAndLiabilitiesMeasuredAtFairValueOnRecurringBasisDetails" roleURI="http://genelux.com/role/ScheduleOfFinancialAssetsAndLiabilitiesMeasuredAtFairValueOnRecurringBasisDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfAvailableForSaleInvestmentsDetails" roleURI="http://genelux.com/role/ScheduleOfAvailableForSaleInvestmentsDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfPropertyAndEquipmentDetails" roleURI="http://genelux.com/role/ScheduleOfPropertyAndEquipmentDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfAccruedExpenseDetails" roleURI="http://genelux.com/role/ScheduleOfAccruedExpenseDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#BalanceSheetAccountsDetailsNarrative" roleURI="http://genelux.com/role/BalanceSheetAccountsDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#LeasesDetailsNarrative" roleURI="http://genelux.com/role/LeasesDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfComponentsOfLeaseAssetsAndLiabilitiesDetails" roleURI="http://genelux.com/role/ScheduleOfComponentsOfLeaseAssetsAndLiabilitiesDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfLeaseCostDetails" roleURI="http://genelux.com/role/ScheduleOfLeaseCostDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfMaturitiesLeaseLiabilitiesDetails" roleURI="http://genelux.com/role/ScheduleOfMaturitiesLeaseLiabilitiesDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfSupplementalCashFlowAndOtherInformationRelatedLeasesDetails" roleURI="http://genelux.com/role/ScheduleOfSupplementalCashFlowAndOtherInformationRelatedLeasesDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfPreferredStockAndCommonStockDetails" roleURI="http://genelux.com/role/ScheduleOfPreferredStockAndCommonStockDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StockholdersEquityDetailsNarrative" roleURI="http://genelux.com/role/StockholdersEquityDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfAwardsOutstandingDetails" roleURI="http://genelux.com/role/ScheduleOfAwardsOutstandingDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfStock-basedCompensationExpensesDetails" roleURI="http://genelux.com/role/ScheduleOfStock-basedCompensationExpensesDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfRestrictedStockUnitsActivityDetails" roleURI="http://genelux.com/role/ScheduleOfRestrictedStockUnitsActivityDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfAssumptionsUsedForOptionsGrantedDetails" roleURI="http://genelux.com/role/ScheduleOfAssumptionsUsedForOptionsGrantedDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfStockOptionActivityDetails" roleURI="http://genelux.com/role/ScheduleOfStockOptionActivityDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfWarrantsActivitiesDetails" roleURI="http://genelux.com/role/ScheduleOfWarrantsActivitiesDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#StockBasedCompensationDetailsNarrative" roleURI="http://genelux.com/role/StockBasedCompensationDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfPotentiallyDilutiveSecuritiesDetails" roleURI="http://genelux.com/role/ScheduleOfPotentiallyDilutiveSecuritiesDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#ScheduleOfSegmentInformationDetails" roleURI="http://genelux.com/role/ScheduleOfSegmentInformationDetails" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SegmentInformationDetailsNarrative" roleURI="http://genelux.com/role/SegmentInformationDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="gnlx-20260630.xsd#SubsequentEventsDetailsNarrative" roleURI="http://genelux.com/role/SubsequentEventsDetailsNarrative" />
    <link:roleRef xlink:type="simple" xlink:href="https://xbrl.sec.gov/ecd/2026/ecd-2026.xsd#PvpExecutiveCategoriesOnly" roleURI="http://xbrl.sec.gov/ecd/role/PvpExecutiveCategoriesOnly" />
    <link:roleRef xlink:type="simple" xlink:href="https://xbrl.sec.gov/ecd/2026/ecd-2026.xsd#PvpAdjustmentsOnly" roleURI="http://xbrl.sec.gov/ecd/role/PvpAdjustmentsOnly" />
    <link:roleRef xlink:type="simple" xlink:href="https://xbrl.sec.gov/ecd/2026/ecd-2026.xsd#IndividualsOnly" roleURI="http://xbrl.sec.gov/ecd/role/IndividualsOnly" />
    <link:roleRef xlink:type="simple" xlink:href="https://xbrl.sec.gov/ecd/2026/ecd-2026.xsd#AwardTypeOnly" roleURI="http://xbrl.sec.gov/ecd/AwardTypeOnly" />
    <link:roleRef xlink:type="simple" xlink:href="https://xbrl.sec.gov/ecd/2026/ecd-2026.xsd#TradingArrangmentsOnly" roleURI="http://xbrl.sec.gov/ecd/role/TradingArrangementsOnly" />
    <link:arcroleRef xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#hypercube-dimension" arcroleURI="http://xbrl.org/int/dim/arcrole/hypercube-dimension" />
    <link:arcroleRef xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#dimension-domain" arcroleURI="http://xbrl.org/int/dim/arcrole/dimension-domain" />
    <link:arcroleRef xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#domain-member" arcroleURI="http://xbrl.org/int/dim/arcrole/domain-member" />
    <link:arcroleRef xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#all" arcroleURI="http://xbrl.org/int/dim/arcrole/all" />
    <link:arcroleRef xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#notAll" arcroleURI="http://xbrl.org/int/dim/arcrole/notAll" />
    <link:arcroleRef xlink:type="simple" xlink:href="http://www.xbrl.org/2005/xbrldt-2005.xsd#dimension-default" arcroleURI="http://xbrl.org/int/dim/arcrole/dimension-default" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/Cover" xlink:title="00000001 - Document - Cover" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BalanceSheets" xlink:title="00000002 - Statement - Condensed Balance Sheets" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BalanceSheetsParenthetical" xlink:title="00000003 - Statement - Condensed Balance Sheets (Parenthetical)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StatementsOfOperationsAndComprehensiveLoss" xlink:title="00000004 - Statement - Condensed Statements of Operations and Comprehensive Loss (Unaudited)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StatementsOfStockholdersEquity" xlink:title="00000005 - Statement - Condensed Statements of Stockholders&apos; Equity (Unaudited)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementLineItems" xlink:label="loc_us-gaapStatementLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementTable" xlink:label="loc_us-gaapStatementTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStatementTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementEquityComponentsAxis" xlink:label="loc_us-gaapStatementEquityComponentsAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapStatementTable" xlink:to="loc_us-gaapStatementEquityComponentsAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EquityComponentDomain" xlink:label="loc_us-gaapEquityComponentDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapStatementEquityComponentsAxis" xlink:to="loc_us-gaapEquityComponentDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EquityComponentDomain" xlink:label="loc_us-gaapEquityComponentDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementEquityComponentsAxis" xlink:to="loc_us-gaapEquityComponentDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockMember" xlink:label="loc_us-gaapCommonStockMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapCommonStockMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TreasuryStockCommonMember" xlink:label="loc_us-gaapTreasuryStockCommonMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapTreasuryStockCommonMember_10" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AdditionalPaidInCapitalMember" xlink:label="loc_us-gaapAdditionalPaidInCapitalMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapAdditionalPaidInCapitalMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AccumulatedOtherComprehensiveIncomeMember" xlink:label="loc_us-gaapAccumulatedOtherComprehensiveIncomeMember_30" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapAccumulatedOtherComprehensiveIncomeMember_30" xlink:type="arc" order="31" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RetainedEarningsMember" xlink:label="loc_us-gaapRetainedEarningsMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapRetainedEarningsMember_40" xlink:type="arc" order="41" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockholdersEquity" xlink:label="loc_us-gaapStockholdersEquity_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockholdersEquity_70" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SharesOutstanding" xlink:label="loc_us-gaapSharesOutstanding_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapSharesOutstanding_70" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TreasuryStockCommonShares" xlink:label="loc_us-gaapTreasuryStockCommonShares_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapTreasuryStockCommonShares_70" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AdjustmentsToAdditionalPaidInCapitalStockIssuedIssuanceCosts" xlink:label="loc_us-gaapAdjustmentsToAdditionalPaidInCapitalStockIssuedIssuanceCosts_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapAdjustmentsToAdditionalPaidInCapitalStockIssuedIssuanceCosts_70" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_OtherComprehensiveIncomeUnrealizedHoldingGainLossOnSecuritiesArisingDuringPeriodNetOfTax" xlink:label="loc_us-gaapOtherComprehensiveIncomeUnrealizedHoldingGainLossOnSecuritiesArisingDuringPeriodNetOfTax_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapOtherComprehensiveIncomeUnrealizedHoldingGainLossOnSecuritiesArisingDuringPeriodNetOfTax_70" xlink:type="arc" order="5" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodValueRestrictedStockAwardNetOfForfeitures" xlink:label="loc_us-gaapStockIssuedDuringPeriodValueRestrictedStockAwardNetOfForfeitures_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodValueRestrictedStockAwardNetOfForfeitures_70" xlink:type="arc" order="6" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesRestrictedStockAwardNetOfForfeitures" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesRestrictedStockAwardNetOfForfeitures_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesRestrictedStockAwardNetOfForfeitures_70" xlink:type="arc" order="7" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_AdjustmentsToAdditionalPaidInCapitalCostOfStockOptionModificationsAndRepricing" xlink:label="loc_GNLXAdjustmentsToAdditionalPaidInCapitalCostOfStockOptionModificationsAndRepricing_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_GNLXAdjustmentsToAdditionalPaidInCapitalCostOfStockOptionModificationsAndRepricing_70" xlink:type="arc" order="8" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodValueNewIssues" xlink:label="loc_us-gaapStockIssuedDuringPeriodValueNewIssues_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodValueNewIssues_70" xlink:type="arc" order="9" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesNewIssues" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesNewIssues_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesNewIssues_70" xlink:type="arc" order="10" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodValueStockOptionsExercised" xlink:label="loc_us-gaapStockIssuedDuringPeriodValueStockOptionsExercised_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodValueStockOptionsExercised_70" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesStockOptionsExercised" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesStockOptionsExercised_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesStockOptionsExercised_70" xlink:type="arc" order="12" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_NetIncomeLoss" xlink:label="loc_us-gaapNetIncomeLoss_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapNetIncomeLoss_70" xlink:type="arc" order="13" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_StockIssuedDuringPeriodValueIssuanceOfCommonSharesForEquityAwardPlans" xlink:label="loc_GNLXStockIssuedDuringPeriodValueIssuanceOfCommonSharesForEquityAwardPlans_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_GNLXStockIssuedDuringPeriodValueIssuanceOfCommonSharesForEquityAwardPlans_70" xlink:type="arc" order="14" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_StockIssuedDuringPeriodSharesIssuanceOfCommonSharesForEquityAwardPlans" xlink:label="loc_GNLXStockIssuedDuringPeriodSharesIssuanceOfCommonSharesForEquityAwardPlans_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_GNLXStockIssuedDuringPeriodSharesIssuanceOfCommonSharesForEquityAwardPlans_70" xlink:type="arc" order="15" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StatementsOfCashFlows" xlink:title="00000006 - Statement - Condensed Statements of Cash Flows (Unaudited)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BasisOfPresentation" xlink:title="999012 - Disclosure - BASIS OF PRESENTATION" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SummaryOfSignificantAccountingPolicies" xlink:title="999013 - Disclosure - SUMMARY OF SIGNIFICANT ACCOUNTING POLICIES" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/LicenseAgreements" xlink:title="999014 - Disclosure - LICENSE AGREEMENTS" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/FairValueMeasurements" xlink:title="999015 - Disclosure - FAIR VALUE MEASUREMENTS" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/Investments" xlink:title="999016 - Disclosure - INVESTMENTS" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BalanceSheetAccounts" xlink:title="999017 - Disclosure - BALANCE SHEET ACCOUNTS" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/Leases" xlink:title="999018 - Disclosure - LEASES" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/CommitmentsAndContingencies" xlink:title="999019 - Disclosure - COMMITMENTS AND CONTINGENCIES" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StockholdersEquity" xlink:title="999020 - Disclosure - STOCKHOLDERS&#8217; EQUITY" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StockBasedCompensation" xlink:title="999021 - Disclosure - STOCK BASED COMPENSATION" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/NetLossPerShare" xlink:title="999022 - Disclosure - NET LOSS PER SHARE" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SegmentInformation" xlink:title="999023 - Disclosure - SEGMENT INFORMATION" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SubsequentEvents" xlink:title="999024 - Disclosure - SUBSEQUENT EVENTS" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SummaryOfSignificantAccountingPoliciesPolicies" xlink:title="999025 - Disclosure - SUMMARY OF SIGNIFICANT ACCOUNTING POLICIES (Policies)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/FairValueMeasurementsTables" xlink:title="999026 - Disclosure - FAIR VALUE MEASUREMENTS (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/InvestmentsTables" xlink:title="999027 - Disclosure - INVESTMENTS (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BalanceSheetAccountsTables" xlink:title="999028 - Disclosure - BALANCE SHEET ACCOUNTS (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/LeasesTables" xlink:title="999029 - Disclosure - LEASES (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StockholdersEquityTables" xlink:title="999030 - Disclosure - STOCKHOLDERS&#8217; EQUITY (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StockBasedCompensationTables" xlink:title="999031 - Disclosure - STOCK BASED COMPENSATION (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/NetLossPerShareTables" xlink:title="999032 - Disclosure - NET LOSS PER SHARE (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SegmentInformationTables" xlink:title="999033 - Disclosure - SEGMENT INFORMATION (Tables)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BasisOfPresentationDetailsNarrative" xlink:title="999034 - Disclosure - BASIS OF PRESENTATION (Details Narrative)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SummaryOfSignificantAccountingPoliciesDetailsNarrative" xlink:title="999035 - Disclosure - SUMMARY OF SIGNIFICANT ACCOUNTING POLICIES (Details Narrative)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CashAndCashEquivalentsLineItems" xlink:label="loc_us-gaapCashAndCashEquivalentsLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfCashAndCashEquivalentsTable" xlink:label="loc_us-gaapScheduleOfCashAndCashEquivalentsTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapCashAndCashEquivalentsLineItems" xlink:to="loc_us-gaapScheduleOfCashAndCashEquivalentsTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CashAndCashEquivalentsAxis" xlink:label="loc_us-gaapCashAndCashEquivalentsAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfCashAndCashEquivalentsTable" xlink:to="loc_us-gaapCashAndCashEquivalentsAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:label="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapCashAndCashEquivalentsAxis" xlink:to="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:label="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapCashAndCashEquivalentsAxis" xlink:to="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_MoneyMarketFundsMember" xlink:label="loc_us-gaapMoneyMarketFundsMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:to="loc_us-gaapMoneyMarketFundsMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CashEquivalentsAtCarryingValue" xlink:label="loc_us-gaapCashEquivalentsAtCarryingValue_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCashAndCashEquivalentsLineItems" xlink:to="loc_us-gaapCashEquivalentsAtCarryingValue_70" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RestrictedCash" xlink:label="loc_us-gaapRestrictedCash_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCashAndCashEquivalentsLineItems" xlink:to="loc_us-gaapRestrictedCash_70" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_UnrealizedGainLossOnInvestments" xlink:label="loc_us-gaapUnrealizedGainLossOnInvestments_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCashAndCashEquivalentsLineItems" xlink:to="loc_us-gaapUnrealizedGainLossOnInvestments_70" xlink:type="arc" order="2" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/LicenseAgreementsDetailsNarrative" xlink:title="999036 - Disclosure - LICENSE AGREEMENTS (Details Narrative)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" xlink:label="loc_us-gaapCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsTable" xlink:label="loc_us-gaapScheduleOfCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" xlink:to="loc_us-gaapScheduleOfCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TypeOfArrangementAxis" xlink:label="loc_us-gaapTypeOfArrangementAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsTable" xlink:to="loc_us-gaapTypeOfArrangementAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_NewsoaraLicenseAgreementMember" xlink:label="loc_GNLXNewsoaraLicenseAgreementMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:to="loc_GNLXNewsoaraLicenseAgreementMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_UpfrontAndMilestonePayment" xlink:label="loc_GNLXUpfrontAndMilestonePayment_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" xlink:to="loc_GNLXUpfrontAndMilestonePayment_20" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_UpfrontPayment" xlink:label="loc_GNLXUpfrontPayment_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" xlink:to="loc_GNLXUpfrontPayment_20" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_MilestonePayment" xlink:label="loc_GNLXMilestonePayment_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" xlink:to="loc_GNLXMilestonePayment_20" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_AdditionalDevelopmentAndCommercialMilestonePayments" xlink:label="loc_GNLXAdditionalDevelopmentAndCommercialMilestonePayments_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapCollaborativeArrangementsAndNoncollaborativeArrangementTransactionsLineItems" xlink:to="loc_GNLXAdditionalDevelopmentAndCommercialMilestonePayments_20" xlink:type="arc" order="3" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfFinancialAssetsAndLiabilitiesMeasuredAtFairValueOnRecurringBasisDetails" xlink:title="999037 - Disclosure - SCHEDULE OF FINANCIAL ASSETS AND LIABILITIES MEASURED AT FAIR VALUE ON A RECURRING BASIS (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueOffBalanceSheetRisksDisclosureInformationLineItems" xlink:label="loc_us-gaapFairValueOffBalanceSheetRisksDisclosureInformationLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfFairValueOffBalanceSheetRisksTable" xlink:label="loc_us-gaapScheduleOfFairValueOffBalanceSheetRisksTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapFairValueOffBalanceSheetRisksDisclosureInformationLineItems" xlink:to="loc_us-gaapScheduleOfFairValueOffBalanceSheetRisksTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueByFairValueHierarchyLevelAxis" xlink:label="loc_us-gaapFairValueByFairValueHierarchyLevelAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfFairValueOffBalanceSheetRisksTable" xlink:to="loc_us-gaapFairValueByFairValueHierarchyLevelAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueMeasurementsFairValueHierarchyDomain" xlink:label="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapFairValueByFairValueHierarchyLevelAxis" xlink:to="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueMeasurementsFairValueHierarchyDomain" xlink:label="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapFairValueByFairValueHierarchyLevelAxis" xlink:to="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueInputsLevel1Member" xlink:label="loc_us-gaapFairValueInputsLevel1Member_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain" xlink:to="loc_us-gaapFairValueInputsLevel1Member_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueInputsLevel2Member" xlink:label="loc_us-gaapFairValueInputsLevel2Member_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain" xlink:to="loc_us-gaapFairValueInputsLevel2Member_10" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FairValueInputsLevel3Member" xlink:label="loc_us-gaapFairValueInputsLevel3Member_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapFairValueMeasurementsFairValueHierarchyDomain" xlink:to="loc_us-gaapFairValueInputsLevel3Member_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CashAndCashEquivalentsAxis" xlink:label="loc_us-gaapCashAndCashEquivalentsAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfFairValueOffBalanceSheetRisksTable" xlink:to="loc_us-gaapCashAndCashEquivalentsAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:label="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapCashAndCashEquivalentsAxis" xlink:to="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:label="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapCashAndCashEquivalentsAxis" xlink:to="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_MoneyMarketFundsMember" xlink:label="loc_us-gaapMoneyMarketFundsMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapRestrictedCashAndCashEquivalentsCashAndCashEquivalentsMember" xlink:to="loc_us-gaapMoneyMarketFundsMember_0" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FinancialInstrumentAxis" xlink:label="loc_us-gaapFinancialInstrumentAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfFairValueOffBalanceSheetRisksTable" xlink:to="loc_us-gaapFinancialInstrumentAxis" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:label="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapFinancialInstrumentAxis" xlink:to="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:label="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapFinancialInstrumentAxis" xlink:to="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_USGovernmentAgenciesDebtSecuritiesMember" xlink:label="loc_us-gaapUSGovernmentAgenciesDebtSecuritiesMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:to="loc_us-gaapUSGovernmentAgenciesDebtSecuritiesMember_40" xlink:type="arc" order="42" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_USTreasurySecuritiesMember" xlink:label="loc_us-gaapUSTreasurySecuritiesMember_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:to="loc_us-gaapUSTreasurySecuritiesMember_80" xlink:type="arc" order="82" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CashAndCashEquivalentsFairValueDisclosure" xlink:label="loc_us-gaapCashAndCashEquivalentsFairValueDisclosure_330" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapFairValueOffBalanceSheetRisksDisclosureInformationLineItems" xlink:to="loc_us-gaapCashAndCashEquivalentsFairValueDisclosure_330" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_MarketableSecurities" xlink:label="loc_us-gaapMarketableSecurities_330" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapFairValueOffBalanceSheetRisksDisclosureInformationLineItems" xlink:to="loc_us-gaapMarketableSecurities_330" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AssetsFairValueDisclosure" xlink:label="loc_us-gaapAssetsFairValueDisclosure_330" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapFairValueOffBalanceSheetRisksDisclosureInformationLineItems" xlink:to="loc_us-gaapAssetsFairValueDisclosure_330" xlink:type="arc" order="2" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfAvailableForSaleInvestmentsDetails" xlink:title="999038 - Disclosure - SCHEDULE OF AVAILABLE FOR SALE INVESTMENTS (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_DebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" xlink:label="loc_us-gaapDebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_DebtSecuritiesHeldToMaturityAllowanceForCreditLossTable" xlink:label="loc_us-gaapDebtSecuritiesHeldToMaturityAllowanceForCreditLossTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapDebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" xlink:to="loc_us-gaapDebtSecuritiesHeldToMaturityAllowanceForCreditLossTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_FinancialInstrumentAxis" xlink:label="loc_us-gaapFinancialInstrumentAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapDebtSecuritiesHeldToMaturityAllowanceForCreditLossTable" xlink:to="loc_us-gaapFinancialInstrumentAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:label="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapFinancialInstrumentAxis" xlink:to="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:label="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapFinancialInstrumentAxis" xlink:to="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_USGovernmentAgenciesDebtSecuritiesMember" xlink:label="loc_us-gaapUSGovernmentAgenciesDebtSecuritiesMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:to="loc_us-gaapUSGovernmentAgenciesDebtSecuritiesMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_USTreasurySecuritiesMember" xlink:label="loc_us-gaapUSTreasurySecuritiesMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapTransfersAndServicingOfFinancialInstrumentsTypesOfFinancialInstrumentsDomain" xlink:to="loc_us-gaapUSTreasurySecuritiesMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AvailableForSaleDebtSecuritiesAmortizedCostBasis" xlink:label="loc_us-gaapAvailableForSaleDebtSecuritiesAmortizedCostBasis_130" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapDebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" xlink:to="loc_us-gaapAvailableForSaleDebtSecuritiesAmortizedCostBasis_130" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AvailableForSaleDebtSecuritiesGrossUnrealizedGain" xlink:label="loc_us-gaapAvailableForSaleDebtSecuritiesGrossUnrealizedGain_130" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapDebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" xlink:to="loc_us-gaapAvailableForSaleDebtSecuritiesGrossUnrealizedGain_130" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AvailableForSaleDebtSecuritiesGrossUnrealizedLoss" xlink:label="loc_us-gaapAvailableForSaleDebtSecuritiesGrossUnrealizedLoss_130" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapDebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" xlink:to="loc_us-gaapAvailableForSaleDebtSecuritiesGrossUnrealizedLoss_130" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_DebtSecuritiesTradingAndAvailableForSale" xlink:label="loc_us-gaapDebtSecuritiesTradingAndAvailableForSale_130" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapDebtSecuritiesHeldtomaturityAllowanceForCreditLossLineItems" xlink:to="loc_us-gaapDebtSecuritiesTradingAndAvailableForSale_130" xlink:type="arc" order="3" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfPropertyAndEquipmentDetails" xlink:title="999039 - Disclosure - SCHEDULE OF PROPERTY AND EQUIPMENT (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentLineItems" xlink:label="loc_us-gaapPropertyPlantAndEquipmentLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfPropertyPlantAndEquipmentTable" xlink:label="loc_us-gaapScheduleOfPropertyPlantAndEquipmentTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapPropertyPlantAndEquipmentLineItems" xlink:to="loc_us-gaapScheduleOfPropertyPlantAndEquipmentTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentByTypeAxis" xlink:label="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfPropertyPlantAndEquipmentTable" xlink:to="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain" xlink:label="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" xlink:to="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain" xlink:label="loc_us-gaapPropertyPlantAndEquipmentTypeDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" xlink:to="loc_us-gaapPropertyPlantAndEquipmentTypeDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_FurnitureAndOfficeEquipmentMember" xlink:label="loc_GNLXFurnitureAndOfficeEquipmentMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_GNLXFurnitureAndOfficeEquipmentMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_LaboratoryEquipmentMember" xlink:label="loc_GNLXLaboratoryEquipmentMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_GNLXLaboratoryEquipmentMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ComputerEquipmentMember" xlink:label="loc_us-gaapComputerEquipmentMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_us-gaapComputerEquipmentMember_40" xlink:type="arc" order="41" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_LeaseholdImprovementsMember" xlink:label="loc_us-gaapLeaseholdImprovementsMember_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_us-gaapLeaseholdImprovementsMember_60" xlink:type="arc" order="61" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ManufacturingEquipmentMember" xlink:label="loc_GNLXManufacturingEquipmentMember_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_GNLXManufacturingEquipmentMember_80" xlink:type="arc" order="81" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ConstructionInProgressMember" xlink:label="loc_us-gaapConstructionInProgressMember_100" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_us-gaapConstructionInProgressMember_100" xlink:type="arc" order="101" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentGross" xlink:label="loc_us-gaapPropertyPlantAndEquipmentGross_150" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentLineItems" xlink:to="loc_us-gaapPropertyPlantAndEquipmentGross_150" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment" xlink:label="loc_us-gaapAccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment_150" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentLineItems" xlink:to="loc_us-gaapAccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment_150" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentNet" xlink:label="loc_us-gaapPropertyPlantAndEquipmentNet_150" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentLineItems" xlink:to="loc_us-gaapPropertyPlantAndEquipmentNet_150" xlink:type="arc" order="2" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfAccruedExpenseDetails" xlink:title="999040 - Disclosure - SCHEDULE OF ACCRUED EXPENSE (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/BalanceSheetAccountsDetailsNarrative" xlink:title="999041 - Disclosure - BALANCE SHEET ACCOUNTS (Details Narrative)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/LeasesDetailsNarrative" xlink:title="999042 - Disclosure - LEASES (Details Narrative)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementLineItems" xlink:label="loc_us-gaapStatementLineItems" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_LeasesTable" xlink:label="loc_GNLXLeasesTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_GNLXLeasesTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_StatementGeographicalAxis" xlink:label="loc_srtStatementGeographicalAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_GNLXLeasesTable" xlink:to="loc_srtStatementGeographicalAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_SegmentGeographicalDomain" xlink:label="loc_srtSegmentGeographicalDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srtStatementGeographicalAxis" xlink:to="loc_srtSegmentGeographicalDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_SegmentGeographicalDomain" xlink:label="loc_srtSegmentGeographicalDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_srtStatementGeographicalAxis" xlink:to="loc_srtSegmentGeographicalDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_WestlakeVillageMember" xlink:label="loc_GNLXWestlakeVillageMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtSegmentGeographicalDomain" xlink:to="loc_GNLXWestlakeVillageMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_SanDiegoMember" xlink:label="loc_GNLXSanDiegoMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtSegmentGeographicalDomain" xlink:to="loc_GNLXSanDiegoMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentByTypeAxis" xlink:label="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_GNLXLeasesTable" xlink:to="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain" xlink:label="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" xlink:to="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PropertyPlantAndEquipmentTypeDomain" xlink:label="loc_us-gaapPropertyPlantAndEquipmentTypeDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapPropertyPlantAndEquipmentByTypeAxis" xlink:to="loc_us-gaapPropertyPlantAndEquipmentTypeDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ResearchAndDevelopmentLaboratoryMember" xlink:label="loc_GNLXResearchAndDevelopmentLaboratoryMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_GNLXResearchAndDevelopmentLaboratoryMember_20" xlink:type="arc" order="22" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ManufacturingSpaceMember" xlink:label="loc_GNLXManufacturingSpaceMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPropertyPlantAndEquipmentTypeDomain" xlink:to="loc_GNLXManufacturingSpaceMember_40" xlink:type="arc" order="42" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TypeOfArrangementAxis" xlink:label="loc_us-gaapTypeOfArrangementAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_GNLXLeasesTable" xlink:to="loc_us-gaapTypeOfArrangementAxis" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_EquipmentAgreementsMember" xlink:label="loc_GNLXEquipmentAgreementsMember_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:to="loc_GNLXEquipmentAgreementsMember_70" xlink:type="arc" order="71" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_RangeAxis" xlink:label="loc_srtRangeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_GNLXLeasesTable" xlink:to="loc_srtRangeAxis" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_RangeMember" xlink:label="loc_srtRangeMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srtRangeAxis" xlink:to="loc_srtRangeMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_RangeMember" xlink:label="loc_srtRangeMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_srtRangeAxis" xlink:to="loc_srtRangeMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_MaximumMember" xlink:label="loc_srtMaximumMember_90" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtRangeMember" xlink:to="loc_srtMaximumMember_90" xlink:type="arc" order="92" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AreaOfLand" xlink:label="loc_us-gaapAreaOfLand_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapAreaOfLand_120" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_LeaseExpirationDate1" xlink:label="loc_us-gaapLeaseExpirationDate1_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapLeaseExpirationDate1_120" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_LesseeOperatingLeaseOptionToExtend" xlink:label="loc_us-gaapLesseeOperatingLeaseOptionToExtend_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapLesseeOperatingLeaseOptionToExtend_120" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_LesseeOperatingLeaseDescription" xlink:label="loc_us-gaapLesseeOperatingLeaseDescription_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapLesseeOperatingLeaseDescription_120" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShortTermLeaseCommitmentAmount" xlink:label="loc_us-gaapShortTermLeaseCommitmentAmount_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapShortTermLeaseCommitmentAmount_120" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PaymentsToAcquireEquipmentOnLease" xlink:label="loc_us-gaapPaymentsToAcquireEquipmentOnLease_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapPaymentsToAcquireEquipmentOnLease_120" xlink:type="arc" order="5" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SecurityDeposit" xlink:label="loc_us-gaapSecurityDeposit_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapSecurityDeposit_120" xlink:type="arc" order="6" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfComponentsOfLeaseAssetsAndLiabilitiesDetails" xlink:title="999043 - Disclosure - SCHEDULE OF COMPONENTS OF LEASE ASSETS AND LIABILITIES (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfLeaseCostDetails" xlink:title="999044 - Disclosure - SCHEDULE OF LEASE COST (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementLineItems" xlink:label="loc_us-gaapStatementLineItems" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_LeasesCostTable" xlink:label="loc_GNLXLeasesCostTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_GNLXLeasesCostTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_StatementScenarioAxis" xlink:label="loc_srtStatementScenarioAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_GNLXLeasesCostTable" xlink:to="loc_srtStatementScenarioAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_ScenarioUnspecifiedDomain" xlink:label="loc_srtScenarioUnspecifiedDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srtStatementScenarioAxis" xlink:to="loc_srtScenarioUnspecifiedDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_ScenarioUnspecifiedDomain" xlink:label="loc_srtScenarioUnspecifiedDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_srtStatementScenarioAxis" xlink:to="loc_srtScenarioUnspecifiedDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ResearchAndDevelopmentExpensesMember" xlink:label="loc_GNLXResearchAndDevelopmentExpensesMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtScenarioUnspecifiedDomain" xlink:to="loc_GNLXResearchAndDevelopmentExpensesMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_GeneralAndAdministrativeExpensesMember" xlink:label="loc_GNLXGeneralAndAdministrativeExpensesMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtScenarioUnspecifiedDomain" xlink:to="loc_GNLXGeneralAndAdministrativeExpensesMember_40" xlink:type="arc" order="41" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_LeaseCost" xlink:label="loc_us-gaapLeaseCost_90" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapStatementLineItems" xlink:to="loc_us-gaapLeaseCost_90" xlink:type="arc" order="0" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfMaturitiesLeaseLiabilitiesDetails" xlink:title="999045 - Disclosure - SCHEDULE OF MATURITIES LEASE LIABILITIES (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfSupplementalCashFlowAndOtherInformationRelatedLeasesDetails" xlink:title="999046 - Disclosure - SCHEDULE OF SUPPLEMENTAL CASH FLOW AND OTHER INFORMATION RELATED LEASES (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfPreferredStockAndCommonStockDetails" xlink:title="999047 - Disclosure - SCHEDULE OF PREFERRED STOCK AND COMMON STOCK (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StockholdersEquityDetailsNarrative" xlink:title="999048 - Disclosure - STOCKHOLDERS&#8217; EQUITY (Details Narrative)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AccumulatedOtherComprehensiveIncomeLossLineItems" xlink:label="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AccumulatedOtherComprehensiveIncomeLossTable" xlink:label="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementEquityComponentsAxis" xlink:label="loc_us-gaapStatementEquityComponentsAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossTable" xlink:to="loc_us-gaapStatementEquityComponentsAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EquityComponentDomain" xlink:label="loc_us-gaapEquityComponentDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapStatementEquityComponentsAxis" xlink:to="loc_us-gaapEquityComponentDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EquityComponentDomain" xlink:label="loc_us-gaapEquityComponentDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementEquityComponentsAxis" xlink:to="loc_us-gaapEquityComponentDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockMember" xlink:label="loc_us-gaapCommonStockMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapCommonStockMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TypeOfArrangementAxis" xlink:label="loc_us-gaapTypeOfArrangementAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossTable" xlink:to="loc_us-gaapTypeOfArrangementAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ATMAgreementMember" xlink:label="loc_GNLXATMAgreementMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:to="loc_GNLXATMAgreementMember_40" xlink:type="arc" order="41" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockSharesAuthorized" xlink:label="loc_us-gaapCommonStockSharesAuthorized_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapCommonStockSharesAuthorized_60" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockSharesIssued" xlink:label="loc_us-gaapCommonStockSharesIssued_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapCommonStockSharesIssued_60" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockSharesOutstanding" xlink:label="loc_us-gaapCommonStockSharesOutstanding_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapCommonStockSharesOutstanding_60" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesNewIssues" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesNewIssues_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesNewIssues_60" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SharePrice" xlink:label="loc_us-gaapSharePrice_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapSharePrice_60" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ProceedsFromIssuanceInitialPublicOffering" xlink:label="loc_us-gaapProceedsFromIssuanceInitialPublicOffering_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapProceedsFromIssuanceInitialPublicOffering_60" xlink:type="arc" order="5" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ProceedsFromCommissionsReceived" xlink:label="loc_us-gaapProceedsFromCommissionsReceived_60" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAccumulatedOtherComprehensiveIncomeLossLineItems" xlink:to="loc_us-gaapProceedsFromCommissionsReceived_60" xlink:type="arc" order="6" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfAwardsOutstandingDetails" xlink:title="999049 - Disclosure - SCHEDULE OF AWARDS OUTSTANDING (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:label="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PlanNameAxis" xlink:label="loc_us-gaapPlanNameAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_us-gaapPlanNameAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PlanNameDomain" xlink:label="loc_us-gaapPlanNameDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapPlanNameAxis" xlink:to="loc_us-gaapPlanNameDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PlanNameDomain" xlink:label="loc_us-gaapPlanNameDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapPlanNameAxis" xlink:to="loc_us-gaapPlanNameDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandNinePlanMember" xlink:label="loc_GNLXTwoThousandNinePlanMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandNinePlanMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandNineteenPlanMember" xlink:label="loc_GNLXTwoThousandNineteenPlanMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandNineteenPlanMember_10" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandTwentyTwoPlanMember" xlink:label="loc_GNLXTwoThousandTwentyTwoPlanMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandTwentyTwoPlanMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_InducementPlanMember" xlink:label="loc_GNLXInducementPlanMember_30" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXInducementPlanMember_30" xlink:type="arc" order="31" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AwardTypeAxis" xlink:label="loc_us-gaapAwardTypeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_us-gaapAwardTypeAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapAwardTypeAxis" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapAwardTypeAxis" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EmployeeStockOptionMember" xlink:label="loc_us-gaapEmployeeStockOptionMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_us-gaapEmployeeStockOptionMember_0" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_RestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember" xlink:label="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember_50" xlink:type="arc" order="52" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardsOutstandingNumber" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardsOutstandingNumber_160" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardsOutstandingNumber_160" xlink:type="arc" order="0" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfStock-basedCompensationExpensesDetails" xlink:title="999050 - Disclosure - SCHEDULE OF STOCK-BASED COMPENSATION EXPENSES (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:label="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_StatementScenarioAxis" xlink:label="loc_srtStatementScenarioAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_srtStatementScenarioAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_ScenarioUnspecifiedDomain" xlink:label="loc_srtScenarioUnspecifiedDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srtStatementScenarioAxis" xlink:to="loc_srtScenarioUnspecifiedDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_ScenarioUnspecifiedDomain" xlink:label="loc_srtScenarioUnspecifiedDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_srtStatementScenarioAxis" xlink:to="loc_srtScenarioUnspecifiedDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_GeneralAndAdministrativeExpensesMember" xlink:label="loc_GNLXGeneralAndAdministrativeExpensesMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtScenarioUnspecifiedDomain" xlink:to="loc_GNLXGeneralAndAdministrativeExpensesMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ResearchAndDevelopmentExpensesMember" xlink:label="loc_GNLXResearchAndDevelopmentExpensesMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtScenarioUnspecifiedDomain" xlink:to="loc_GNLXResearchAndDevelopmentExpensesMember_40" xlink:type="arc" order="41" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AwardTypeAxis" xlink:label="loc_us-gaapAwardTypeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_us-gaapAwardTypeAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapAwardTypeAxis" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapAwardTypeAxis" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EmployeeStockOptionMember" xlink:label="loc_us-gaapEmployeeStockOptionMember_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_us-gaapEmployeeStockOptionMember_120" xlink:type="arc" order="121" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_RestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember" xlink:label="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember_160" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember_160" xlink:type="arc" order="161" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_EmployeeStockPurchasePlanMember" xlink:label="loc_GNLXEmployeeStockPurchasePlanMember_200" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXEmployeeStockPurchasePlanMember_200" xlink:type="arc" order="201" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AllocatedShareBasedCompensationExpense" xlink:label="loc_us-gaapAllocatedShareBasedCompensationExpense_250" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapAllocatedShareBasedCompensationExpense_250" xlink:type="arc" order="0" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfRestrictedStockUnitsActivityDetails" xlink:title="999051 - Disclosure - SCHEDULE OF RESTRICTED STOCK UNITS ACTIVITY (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfAssumptionsUsedForOptionsGrantedDetails" xlink:title="999052 - Disclosure - SCHEDULE OF ASSUMPTIONS USED FOR OPTIONS GRANTED (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfStockOptionActivityDetails" xlink:title="999053 - Disclosure - SCHEDULE OF STOCK OPTION ACTIVITY (Details)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfWarrantsActivitiesDetails" xlink:title="999054 - Disclosure - SCHEDULE OF WARRANTS ACTIVITIES (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:label="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_RangeAxis" xlink:label="loc_srtRangeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_srtRangeAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_RangeMember" xlink:label="loc_srtRangeMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srtRangeAxis" xlink:to="loc_srtRangeMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_RangeMember" xlink:label="loc_srtRangeMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_srtRangeAxis" xlink:to="loc_srtRangeMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_MinimumMember" xlink:label="loc_srtMinimumMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtRangeMember" xlink:to="loc_srtMinimumMember_10" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_MaximumMember" xlink:label="loc_srtMaximumMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtRangeMember" xlink:to="loc_srtMaximumMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingNumber" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingNumber_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingNumber_80" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingExercisePriceRange" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingExercisePriceRange_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingExercisePriceRange_80" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingWeightedAverageExercisePrice" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingWeightedAverageExercisePrice_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsOutstandingWeightedAverageExercisePrice_80" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsGranted" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsGranted_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsGranted_80" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsGrantedExercisePriceRange" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsGrantedExercisePriceRange_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsGrantedExercisePriceRange_80" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsIssuedWeightedAverageExercisePrice" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsIssuedWeightedAverageExercisePrice_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsIssuedWeightedAverageExercisePrice_80" xlink:type="arc" order="5" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitures" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitures_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitures_80" xlink:type="arc" order="6" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitedExercisePriceRange" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitedExercisePriceRange_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitedExercisePriceRange_80" xlink:type="arc" order="7" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitedWeightedAverageExercisePrice" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitedWeightedAverageExercisePrice_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsForfeitedWeightedAverageExercisePrice_80" xlink:type="arc" order="8" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercised" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercised_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercised_80" xlink:type="arc" order="9" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisedExercisePriceRange" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisedExercisePriceRange_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisedExercisePriceRange_80" xlink:type="arc" order="10" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisedWeightedAverageExercisePrice" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisedWeightedAverageExercisePrice_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisedWeightedAverageExercisePrice_80" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpirations" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpirations_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpirations_80" xlink:type="arc" order="12" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpiredExercisePriceRange" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpiredExercisePriceRange_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpiredExercisePriceRange_80" xlink:type="arc" order="13" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpiredWeightedAverageExercisePrice" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpiredWeightedAverageExercisePrice_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExpiredWeightedAverageExercisePrice_80" xlink:type="arc" order="14" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableNumber" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableNumber_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableNumber_80" xlink:type="arc" order="18" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableExercisePriceRange" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableExercisePriceRange_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableExercisePriceRange_80" xlink:type="arc" order="19" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableWeightedAverageExercisePrice" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableWeightedAverageExercisePrice_80" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionEquityInstrumentsExercisableWeightedAverageExercisePrice_80" xlink:type="arc" order="20" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/StockBasedCompensationDetailsNarrative" xlink:title="999055 - Disclosure - STOCK BASED COMPENSATION (Details Narrative)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:label="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PlanNameAxis" xlink:label="loc_us-gaapPlanNameAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_us-gaapPlanNameAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PlanNameDomain" xlink:label="loc_us-gaapPlanNameDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapPlanNameAxis" xlink:to="loc_us-gaapPlanNameDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_PlanNameDomain" xlink:label="loc_us-gaapPlanNameDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapPlanNameAxis" xlink:to="loc_us-gaapPlanNameDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandNinePlanMember" xlink:label="loc_GNLXTwoThousandNinePlanMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandNinePlanMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandNineteenPlanMember" xlink:label="loc_GNLXTwoThousandNineteenPlanMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandNineteenPlanMember_10" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandTwentyTwoPlanMember" xlink:label="loc_GNLXTwoThousandTwentyTwoPlanMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandTwentyTwoPlanMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandTwentyThreeInducementPlanMember" xlink:label="loc_GNLXTwoThousandTwentyThreeInducementPlanMember_70" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandTwentyThreeInducementPlanMember_70" xlink:type="arc" order="71" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_InducementPlanMember" xlink:label="loc_GNLXInducementPlanMember_180" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXInducementPlanMember_180" xlink:type="arc" order="181" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_TwoThousandTwentyTwoEmployeeStockPurchasePlanMember" xlink:label="loc_GNLXTwoThousandTwentyTwoEmployeeStockPurchasePlanMember_240" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapPlanNameDomain" xlink:to="loc_GNLXTwoThousandTwentyTwoEmployeeStockPurchasePlanMember_240" xlink:type="arc" order="241" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementEquityComponentsAxis" xlink:label="loc_us-gaapStatementEquityComponentsAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_us-gaapStatementEquityComponentsAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EquityComponentDomain" xlink:label="loc_us-gaapEquityComponentDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapStatementEquityComponentsAxis" xlink:to="loc_us-gaapEquityComponentDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EquityComponentDomain" xlink:label="loc_us-gaapEquityComponentDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementEquityComponentsAxis" xlink:to="loc_us-gaapEquityComponentDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockMember" xlink:label="loc_us-gaapCommonStockMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapEquityComponentDomain" xlink:to="loc_us-gaapCommonStockMember_0" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AwardTypeAxis" xlink:label="loc_us-gaapAwardTypeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_us-gaapAwardTypeAxis" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapAwardTypeAxis" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapAwardTypeAxis" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_PerformanceStockUnitsPSUMember" xlink:label="loc_GNLXPerformanceStockUnitsPSUMember_100" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXPerformanceStockUnitsPSUMember_100" xlink:type="arc" order="101" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_RestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember" xlink:label="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember_120" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember_120" xlink:type="arc" order="121" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_TitleOfIndividualAxis" xlink:label="loc_srtTitleOfIndividualAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfShareBasedCompensationArrangementsByShareBasedPaymentAwardTable" xlink:to="loc_srtTitleOfIndividualAxis" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_TitleOfIndividualWithRelationshipToEntityDomain" xlink:label="loc_srtTitleOfIndividualWithRelationshipToEntityDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_srtTitleOfIndividualAxis" xlink:to="loc_srtTitleOfIndividualWithRelationshipToEntityDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/srt/2026/elts/srt-2026.xsd#srt_TitleOfIndividualWithRelationshipToEntityDomain" xlink:label="loc_srtTitleOfIndividualWithRelationshipToEntityDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_srtTitleOfIndividualAxis" xlink:to="loc_srtTitleOfIndividualWithRelationshipToEntityDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_EmployeesDirectorsAndKeyAdvisersMember" xlink:label="loc_GNLXEmployeesDirectorsAndKeyAdvisersMember_210" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_srtTitleOfIndividualWithRelationshipToEntityDomain" xlink:to="loc_GNLXEmployeesDirectorsAndKeyAdvisersMember_210" xlink:type="arc" order="211" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardSharesIssuedInPeriod" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardSharesIssuedInPeriod_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardSharesIssuedInPeriod_280" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_CommonStockCapitalSharesReservedForFutureIssuance" xlink:label="loc_us-gaapCommonStockCapitalSharesReservedForFutureIssuance_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapCommonStockCapitalSharesReservedForFutureIssuance_280" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_NumberOfSharesOfCommonStockPercentage" xlink:label="loc_GNLXNumberOfSharesOfCommonStockPercentage_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXNumberOfSharesOfCommonStockPercentage_280" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNumberOfSharesAvailableForGrant" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNumberOfSharesAvailableForGrant_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardNumberOfSharesAvailableForGrant_280" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesIssuedForServices" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesIssuedForServices_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesIssuedForServices_280" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesNewIssues" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesNewIssues_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesNewIssues_280" xlink:type="arc" order="5" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsGrantsInPeriod" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsGrantsInPeriod_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsGrantsInPeriod_280" xlink:type="arc" order="6" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AdjustmentForAmortization" xlink:label="loc_us-gaapAdjustmentForAmortization_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapAdjustmentForAmortization_280" xlink:type="arc" order="7" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriod" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriod_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardEquityInstrumentsOtherThanOptionsGrantsInPeriod_280" xlink:type="arc" order="8" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensation" xlink:label="loc_us-gaapShareBasedCompensation_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensation_280" xlink:type="arc" order="9" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_UnamortizedDebtIssuanceExpense" xlink:label="loc_us-gaapUnamortizedDebtIssuanceExpense_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapUnamortizedDebtIssuanceExpense_280" xlink:type="arc" order="10" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_FairMarketValueOfSharesPercentageDescription" xlink:label="loc_GNLXFairMarketValueOfSharesPercentageDescription_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXFairMarketValueOfSharesPercentageDescription_280" xlink:type="arc" order="11" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardOptionsExercisesInPeriodWeightedAverageExercisePrice" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardOptionsExercisesInPeriodWeightedAverageExercisePrice_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardOptionsExercisesInPeriodWeightedAverageExercisePrice_280" xlink:type="arc" order="12" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingWeightedAverageExercisePrice" xlink:label="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingWeightedAverageExercisePrice_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingWeightedAverageExercisePrice_280" xlink:type="arc" order="13" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_RepricingCosts" xlink:label="loc_GNLXRepricingCosts_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXRepricingCosts_280" xlink:type="arc" order="14" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationSharesAuthorizedUnderStockOptionPlansExercisePriceRangeLowerRangeLimit" xlink:label="loc_us-gaapShareBasedCompensationSharesAuthorizedUnderStockOptionPlansExercisePriceRangeLowerRangeLimit_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationSharesAuthorizedUnderStockOptionPlansExercisePriceRangeLowerRangeLimit_280" xlink:type="arc" order="15" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationSharesAuthorizedUnderStockOptionPlansExercisePriceRangeUpperRangeLimit" xlink:label="loc_us-gaapShareBasedCompensationSharesAuthorizedUnderStockOptionPlansExercisePriceRangeUpperRangeLimit_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationSharesAuthorizedUnderStockOptionPlansExercisePriceRangeUpperRangeLimit_280" xlink:type="arc" order="16" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardOptionsGrantsInPeriodWeightedAverageExercisePrice" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardOptionsGrantsInPeriodWeightedAverageExercisePrice_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardOptionsGrantsInPeriodWeightedAverageExercisePrice_280" xlink:type="arc" order="17" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockOptionPlanExpense" xlink:label="loc_us-gaapStockOptionPlanExpense_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapStockOptionPlanExpense_280" xlink:type="arc" order="18" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionsOutstandingIntrinsicValue" xlink:label="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionsOutstandingIntrinsicValue_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXShareBasedCompensationArrangementByShareBasedPaymentAwardNonOptionsOutstandingIntrinsicValue_280" xlink:type="arc" order="19" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_EmployeeStockPurchasePlanDescription" xlink:label="loc_GNLXEmployeeStockPurchasePlanDescription_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_GNLXEmployeeStockPurchasePlanDescription_280" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockIssuedDuringPeriodSharesEmployeeStockPurchasePlans" xlink:label="loc_us-gaapStockIssuedDuringPeriodSharesEmployeeStockPurchasePlans_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapStockIssuedDuringPeriodSharesEmployeeStockPurchasePlans_280" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StockRepurchaseProgramRemainingNumberOfSharesAuthorizedToBeRepurchased" xlink:label="loc_us-gaapStockRepurchaseProgramRemainingNumberOfSharesAuthorizedToBeRepurchased_280" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementByShareBasedPaymentAwardLineItems" xlink:to="loc_us-gaapStockRepurchaseProgramRemainingNumberOfSharesAuthorizedToBeRepurchased_280" xlink:type="arc" order="22" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfPotentiallyDilutiveSecuritiesDetails" xlink:title="999056 - Disclosure - SCHEDULE OF POTENTIALLY DILUTIVE SECURITIES (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems" xlink:label="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable" xlink:label="loc_us-gaapScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems" xlink:to="loc_us-gaapScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareByAntidilutiveSecuritiesAxis" xlink:label="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareByAntidilutiveSecuritiesAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareTable" xlink:to="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareByAntidilutiveSecuritiesAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AntidilutiveSecuritiesNameDomain" xlink:label="loc_us-gaapAntidilutiveSecuritiesNameDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareByAntidilutiveSecuritiesAxis" xlink:to="loc_us-gaapAntidilutiveSecuritiesNameDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AntidilutiveSecuritiesNameDomain" xlink:label="loc_us-gaapAntidilutiveSecuritiesNameDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareByAntidilutiveSecuritiesAxis" xlink:to="loc_us-gaapAntidilutiveSecuritiesNameDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EmployeeStockOptionMember" xlink:label="loc_us-gaapEmployeeStockOptionMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAntidilutiveSecuritiesNameDomain" xlink:to="loc_us-gaapEmployeeStockOptionMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_WarrantMember" xlink:label="loc_us-gaapWarrantMember_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAntidilutiveSecuritiesNameDomain" xlink:to="loc_us-gaapWarrantMember_20" xlink:type="arc" order="21" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_RestrictedStockUnitsRSUAndPSUMember" xlink:label="loc_GNLXRestrictedStockUnitsRSUAndPSUMember_40" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAntidilutiveSecuritiesNameDomain" xlink:to="loc_GNLXRestrictedStockUnitsRSUAndPSUMember_40" xlink:type="arc" order="41" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_AntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareAmount" xlink:label="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareAmount_90" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareLineItems" xlink:to="loc_us-gaapAntidilutiveSecuritiesExcludedFromComputationOfEarningsPerShareAmount_90" xlink:type="arc" order="0" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/ScheduleOfSegmentInformationDetails" xlink:title="999057 - Disclosure - SCHEDULE OF SEGMENT INFORMATION (Details)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SegmentReportingInformationLineItems" xlink:label="loc_us-gaapSegmentReportingInformationLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ScheduleOfSegmentReportingInformationBySegmentTable" xlink:label="loc_us-gaapScheduleOfSegmentReportingInformationBySegmentTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapScheduleOfSegmentReportingInformationBySegmentTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_StatementBusinessSegmentsAxis" xlink:label="loc_us-gaapStatementBusinessSegmentsAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapScheduleOfSegmentReportingInformationBySegmentTable" xlink:to="loc_us-gaapStatementBusinessSegmentsAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SegmentDomain" xlink:label="loc_us-gaapSegmentDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapStatementBusinessSegmentsAxis" xlink:to="loc_us-gaapSegmentDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SegmentDomain" xlink:label="loc_us-gaapSegmentDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapStatementBusinessSegmentsAxis" xlink:to="loc_us-gaapSegmentDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_OperatingSegmentMember" xlink:label="loc_GNLXOperatingSegmentMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentDomain" xlink:to="loc_GNLXOperatingSegmentMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_RevenueFromContractWithCustomerExcludingAssessedTax" xlink:label="loc_us-gaapRevenueFromContractWithCustomerExcludingAssessedTax_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapRevenueFromContractWithCustomerExcludingAssessedTax_50" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ResearchAndDevelopmentExcludingSalaries" xlink:label="loc_GNLXResearchAndDevelopmentExcludingSalaries_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_GNLXResearchAndDevelopmentExcludingSalaries_50" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SalariesAndWages" xlink:label="loc_us-gaapSalariesAndWages_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapSalariesAndWages_50" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_InsuranceCommissions" xlink:label="loc_us-gaapInsuranceCommissions_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapInsuranceCommissions_50" xlink:type="arc" order="3" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_EmployeeBenefitsAndShareBasedCompensation" xlink:label="loc_us-gaapEmployeeBenefitsAndShareBasedCompensation_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapEmployeeBenefitsAndShareBasedCompensation_50" xlink:type="arc" order="4" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_OperatingExpenses" xlink:label="loc_us-gaapOperatingExpenses_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapOperatingExpenses_50" xlink:type="arc" order="5" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_OperatingIncomeLoss" xlink:label="loc_us-gaapOperatingIncomeLoss_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapOperatingIncomeLoss_50" xlink:type="arc" order="6" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_OtherIncome" xlink:label="loc_us-gaapOtherIncome_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapOtherIncome_50" xlink:type="arc" order="7" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_NetIncomeLoss" xlink:label="loc_us-gaapNetIncomeLoss_50" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSegmentReportingInformationLineItems" xlink:to="loc_us-gaapNetIncomeLoss_50" xlink:type="arc" order="8" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SegmentInformationDetailsNarrative" xlink:title="999058 - Disclosure - SEGMENT INFORMATION (Details Narrative)" />
    <link:definitionLink xlink:type="extended" xlink:role="http://genelux.com/role/SubsequentEventsDetailsNarrative" xlink:title="999059 - Disclosure - SUBSEQUENT EVENTS (Details Narrative)">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SubsequentEventLineItems" xlink:label="loc_us-gaapSubsequentEventLineItems" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SubsequentEventTable" xlink:label="loc_us-gaapSubsequentEventTable" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/all" xbrldt:contextElement="segment" xlink:from="loc_us-gaapSubsequentEventLineItems" xlink:to="loc_us-gaapSubsequentEventTable" xlink:type="arc" order="10" xbrldt:closed="true" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SubsequentEventTypeAxis" xlink:label="loc_us-gaapSubsequentEventTypeAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapSubsequentEventTable" xlink:to="loc_us-gaapSubsequentEventTypeAxis" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SubsequentEventTypeDomain" xlink:label="loc_us-gaapSubsequentEventTypeDomain" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapSubsequentEventTypeAxis" xlink:to="loc_us-gaapSubsequentEventTypeDomain" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SubsequentEventTypeDomain" xlink:label="loc_us-gaapSubsequentEventTypeDomain_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapSubsequentEventTypeAxis" xlink:to="loc_us-gaapSubsequentEventTypeDomain_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SubsequentEventMember" xlink:label="loc_us-gaapSubsequentEventMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSubsequentEventTypeDomain" xlink:to="loc_us-gaapSubsequentEventMember_0" xlink:type="arc" order="1" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_TypeOfArrangementAxis" xlink:label="loc_us-gaapTypeOfArrangementAxis" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/hypercube-dimension" xlink:from="loc_us-gaapSubsequentEventTable" xlink:to="loc_us-gaapTypeOfArrangementAxis" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-domain" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:type="arc" order="20" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ArrangementsAndNonarrangementTransactionsMember" xlink:label="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/dimension-default" xbrldt:contextElement="segment" xlink:from="loc_us-gaapTypeOfArrangementAxis" xlink:to="loc_us-gaapArrangementsAndNonarrangementTransactionsMember_10" xlink:type="arc" order="30" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_ATMAgreementMember" xlink:label="loc_GNLXATMAgreementMember_0" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapArrangementsAndNonarrangementTransactionsMember" xlink:to="loc_GNLXATMAgreementMember_0" xlink:type="arc" order="2" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_SaleOfStockNumberOfSharesIssuedInTransaction" xlink:label="loc_us-gaapSaleOfStockNumberOfSharesIssuedInTransaction_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSubsequentEventLineItems" xlink:to="loc_us-gaapSaleOfStockNumberOfSharesIssuedInTransaction_20" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ProceedsFromIssuanceOfCommonStock" xlink:label="loc_us-gaapProceedsFromIssuanceOfCommonStock_20" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapSubsequentEventLineItems" xlink:to="loc_us-gaapProceedsFromIssuanceOfCommonStock_20" xlink:type="arc" order="1" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://xbrl.sec.gov/ecd/role/PvpExecutiveCategoriesOnly" />
    <link:definitionLink xlink:type="extended" xlink:role="http://xbrl.sec.gov/ecd/role/PvpAdjustmentsOnly" />
    <link:definitionLink xlink:type="extended" xlink:role="http://xbrl.sec.gov/ecd/role/IndividualsOnly">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.sec.gov/ecd/2026/ecd-2026.xsd#ecd_AllIndividualsMember" xlink:label="loc_ecdAllIndividualsMember" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_JohnThomasMember" xlink:label="loc_GNLXJohnThomasMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_ecdAllIndividualsMember" xlink:to="loc_GNLXJohnThomasMember" xlink:type="arc" order="0" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://xbrl.sec.gov/ecd/AwardTypeOnly">
      <link:loc xlink:type="locator" xlink:href="https://xbrl.fasb.org/us-gaap/2026/elts/us-gaap-2026.xsd#us-gaap_ShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:label="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_PerformanceStockUnitsPSUMember" xlink:label="loc_GNLXPerformanceStockUnitsPSUMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXPerformanceStockUnitsPSUMember" xlink:type="arc" order="0" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_RestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember" xlink:label="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXRestrictedStockUnitsRSUAndPerformanceStockUnitsPSUsMember" xlink:type="arc" order="10" />
      <link:loc xlink:type="locator" xlink:href="gnlx-20260630.xsd#GNLX_EmployeeStockPurchasePlanMember" xlink:label="loc_GNLXEmployeeStockPurchasePlanMember" />
      <link:definitionArc xlink:arcrole="http://xbrl.org/int/dim/arcrole/domain-member" xlink:from="loc_us-gaapShareBasedCompensationArrangementsByShareBasedPaymentAwardAwardTypeAndPlanNameDomain" xlink:to="loc_GNLXEmployeeStockPurchasePlanMember" xlink:type="arc" order="20" />
    </link:definitionLink>
    <link:definitionLink xlink:type="extended" xlink:role="http://xbrl.sec.gov/ecd/role/TradingArrangementsOnly" />
</link:linkbase>
