<?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>0002145406</rptOwnerCik>
            <rptOwnerName>Richton Kathleen P.</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>SVP, Finance and Treasurer</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <nonDerivativeTable>
        <nonDerivativeHolding>
            <securityTitle>
                <value>Common Shares</value>
            </securityTitle>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>6900</value>
                    <footnoteId id="F1"/>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeHolding>
    </nonDerivativeTable>

    <derivativeTable>
        <derivativeHolding>
            <securityTitle>
                <value>Stock Option (Right to Buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>7.39</value>
            </conversionOrExercisePrice>
            <exerciseDate>
                <footnoteId id="F2"/>
            </exerciseDate>
            <expirationDate>
                <value>2035-01-29</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Shares</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>45000</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeHolding>
        <derivativeHolding>
            <securityTitle>
                <value>Stock Option (Right to Buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>9.53</value>
            </conversionOrExercisePrice>
            <exerciseDate>
                <footnoteId id="F3"/>
            </exerciseDate>
            <expirationDate>
                <value>2036-01-30</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Shares</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>13900</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeHolding>
        <derivativeHolding>
            <securityTitle>
                <value>Stock Option (Right to Buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>1.75</value>
            </conversionOrExercisePrice>
            <exerciseDate>
                <footnoteId id="F4"/>
            </exerciseDate>
            <expirationDate>
                <value>2036-06-16</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Shares</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>105000</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeHolding>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">Consists of 6,900 unvested restricted stock units (RSUs) granted to the Reporting Person on January 30, 2026 pursuant to the Issuer's Amended and Restated 2023 Incentive Equity Plan (the &quot;Plan&quot;), which are scheduled to vest annually in substantially equal amounts for four years, commencing January 15, 2027, subject to the Reporting Person's continued service. Each RSU represents a contingent right to receive one common share of the Issuer.</footnote>
        <footnote id="F2">Non-qualified stock option grant awarded as an inducement award outside of the Plan in accordance with NASDAQ Listing Rule 5635(c)(4). This option vested at 25% on January 27, 2026, with the remaining portion to vest monthly in substantially equal amounts for the following 36 months, subject to the Reporting Person's continued service.</footnote>
        <footnote id="F3">This option vests monthly in substantially equal amounts for 48 months, commencing January 30, 2026, subject to the Reporting Person's continued service.</footnote>
        <footnote id="F4">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</signatureName>
        <signatureDate>2026-07-22</signatureDate>
    </ownerSignature>
</ownershipDocument>
