<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0607</schemaVersion>

    <documentType>3</documentType>

    <periodOfReport>2026-07-16</periodOfReport>

    <noSecuritiesOwned>0</noSecuritiesOwned>

    <issuer>
        <issuerCik>0001980845</issuerCik>
        <issuerName>enGene Therapeutics Inc.</issuerName>
        <issuerTradingSymbol>ENGN</issuerTradingSymbol>
        <issuerForeignTradingSymbol></issuerForeignTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0002014509</rptOwnerCik>
            <rptOwnerName>Chinoporos Constantine</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerNonUSAddressFlag>true</rptOwnerNonUSAddressFlag>
            <rptOwnerStreet1>C/O 4868 RUE LEVY, SUITE 220</rptOwnerStreet1>
            <rptOwnerStreet2></rptOwnerStreet2>
            <rptOwnerCity>SAINT-LAURENT</rptOwnerCity>
            <rptOwnerNonUSStateTerritory></rptOwnerNonUSStateTerritory>
            <rptOwnerCountry>A8</rptOwnerCountry>
            <rptOwnerZipCode>H4R 2P1</rptOwnerZipCode>
            <rptOwnerStateDescription>QUEBEC, CANADA</rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isOfficer>true</isOfficer>
            <officerTitle>Interim Chief Business Officer</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <derivativeTable>
        <derivativeHolding>
            <securityTitle>
                <value>Stock Option (Right to Buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>1.75</value>
            </conversionOrExercisePrice>
            <exerciseDate>
                <footnoteId id="F1"/>
            </exerciseDate>
            <expirationDate>
                <value>2036-06-16</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Shares</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>75000</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeHolding>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">This option vests monthly in substantially equal amounts for 48 months, commencing June 16, 2026, subject to the Reporting Person's continued service.</footnote>
    </footnotes>

    <remarks>Exhibit 24 - Power of Attorney</remarks>

    <ownerSignature>
        <signatureName>/s/ Kathleen P. Richton, as attorney-in-fact for the Reporting Person</signatureName>
        <signatureDate>2026-07-22</signatureDate>
    </ownerSignature>
</ownershipDocument>
