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

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2024-12-23</periodOfReport>

    <issuer>
        <issuerCik>0001623526</issuerCik>
        <issuerName>Stoke Therapeutics, Inc.</issuerName>
        <issuerTradingSymbol>STOK</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001869333</rptOwnerCik>
            <rptOwnerName>Skorpios Trust</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>195 ARCH. MAKARIOS III AVE.</rptOwnerStreet1>
            <rptOwnerStreet2>NEOCLEOUS HOUSE</rptOwnerStreet2>
            <rptOwnerCity>LIMASSOL</rptOwnerCity>
            <rptOwnerState>G4</rptOwnerState>
            <rptOwnerZipCode>3030</rptOwnerZipCode>
            <rptOwnerStateDescription>CYPRUS</rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>0</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>1</isTenPercentOwner>
            <isOther>0</isOther>
        </reportingOwnerRelationship>
    </reportingOwner>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001869057</rptOwnerCik>
            <rptOwnerName>Blue Horizon Enterprise Ltd</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>195 ARCH. MAKARIOS III AVE.</rptOwnerStreet1>
            <rptOwnerStreet2>NEOCLEOUS HOUSE</rptOwnerStreet2>
            <rptOwnerCity>LIMASSOL</rptOwnerCity>
            <rptOwnerState>G4</rptOwnerState>
            <rptOwnerZipCode>3030</rptOwnerZipCode>
            <rptOwnerStateDescription>CYPRUS</rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>0</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>1</isTenPercentOwner>
            <isOther>0</isOther>
        </reportingOwnerRelationship>
    </reportingOwner>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001869027</rptOwnerCik>
            <rptOwnerName>Ezbon International Ltd</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>195 ARCH. MAKARIOS III AVE.</rptOwnerStreet1>
            <rptOwnerStreet2>NEOCLEOUS HOUSE</rptOwnerStreet2>
            <rptOwnerCity>LIMASSOL</rptOwnerCity>
            <rptOwnerState>G4</rptOwnerState>
            <rptOwnerZipCode>3030</rptOwnerZipCode>
            <rptOwnerStateDescription>CYPRUS</rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>0</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>1</isTenPercentOwner>
            <isOther>0</isOther>
        </reportingOwnerRelationship>
    </reportingOwner>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001869331</rptOwnerCik>
            <rptOwnerName>Montrago Trustees Ltd</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>195 ARCH. MAKARIOS III AVE</rptOwnerStreet1>
            <rptOwnerStreet2>NEOCLEOUS HOUSE</rptOwnerStreet2>
            <rptOwnerCity>LIMASSOL</rptOwnerCity>
            <rptOwnerState>G4</rptOwnerState>
            <rptOwnerZipCode>3030</rptOwnerZipCode>
            <rptOwnerStateDescription>CYPRUS</rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>0</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>1</isTenPercentOwner>
            <isOther>0</isOther>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <nonDerivativeTable>
        <nonDerivativeTransaction>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <transactionDate>
                <value>2024-12-23</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>S</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>2000000</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>11.50</value>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>6906181</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>I</value>
                </directOrIndirectOwnership>
                <natureOfOwnership>
                    <value>See Footnote</value>
                    <footnoteId id="F1"/>
                </natureOfOwnership>
            </ownershipNature>
        </nonDerivativeTransaction>
    </nonDerivativeTable>

    <footnotes>
        <footnote id="F1">Blue Horizon Enterprise Ltd. (&quot;Blue Horizon&quot;) is the direct owner of 2,967,621  shares of the issuer's common stock and Ezbon International Limited (&quot;Ezbon&quot;) is the direct owner of 3,938,560 shares of the issuer's common stock. Skorpios Trust (&quot;Skorpios&quot;) is the sole owner of each of Blue Horizon and Ezbon. Montrago Trustees Limited (&quot;Montrago Trustees&quot;) is the corporate trustee of Skorpios Trust. Each of Blue Horizon and Ezbon disclaim beneficial ownership of the shares of the issuer's common stock held by the other, and each of Skorpios Trust and Montrago Trustees disclaim beneficial ownership of the issuer's securities except to the extent of their pecuniary interest therein.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Androulla Papadopoulou, Authorized Signatory on behalf of Skorpios Trust</signatureName>
        <signatureDate>2024-12-27</signatureDate>
    </ownerSignature>

    <ownerSignature>
        <signatureName>/s/ Eleni Constantinou Kyriacou, Authorized Signatory on behalf of Blue Horizon Enterprises Ltd.</signatureName>
        <signatureDate>2024-12-27</signatureDate>
    </ownerSignature>

    <ownerSignature>
        <signatureName>/s/ Eleni Constantinou Kyriacou, Authorized Signatory on behalf of Ezbon International Limited</signatureName>
        <signatureDate>2024-12-27</signatureDate>
    </ownerSignature>

    <ownerSignature>
        <signatureName>/s/ Androulla Papadopoulou, Authorized Signatory on behalf of Montrago Trustees Limited</signatureName>
        <signatureDate>2024-12-27</signatureDate>
    </ownerSignature>
</ownershipDocument>
