<?xml version="1.0"?>
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    <schemaVersion>X0708</schemaVersion>

    <submissionType>D/A</submissionType>

    <testOrLive>LIVE</testOrLive>

    <primaryIssuer>
        <cik>0001932386</cik>
        <entityName>SightStream Biotherapeutics, Inc.</entityName>
        <issuerAddress>
            <street1>266 MAIN ST., BLDG 2, STE 11</street1>
            <city>MEDFIELD</city>
            <stateOrCountry>MA</stateOrCountry>
            <stateOrCountryDescription>MASSACHUSETTS</stateOrCountryDescription>
            <zipCode>02052</zipCode>
        </issuerAddress>
        <issuerPhoneNumber>617-388-2635</issuerPhoneNumber>
        <jurisdictionOfInc>DELAWARE</jurisdictionOfInc>
        <edgarPreviousNameList>
            <previousName>Kera Therapeutics, Inc.</previousName>
        </edgarPreviousNameList>
        <entityType>Corporation</entityType>
        <yearOfInc>
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            <value>2020</value>
        </yearOfInc>
    </primaryIssuer>

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            <relatedPersonName>
                <firstName>Jason</firstName>
                <lastName>Werner</lastName>
            </relatedPersonName>
            <relatedPersonAddress>
                <street1>266 Main St., BLDG 2, STE 11</street1>
                <city>Medfield</city>
                <stateOrCountry>MA</stateOrCountry>
                <stateOrCountryDescription>MASSACHUSETTS</stateOrCountryDescription>
                <zipCode>02052</zipCode>
            </relatedPersonAddress>
            <relatedPersonRelationshipList>
                <relationship>Executive Officer</relationship>
                <relationship>Director</relationship>
            </relatedPersonRelationshipList>
            <relationshipClarification></relationshipClarification>
        </relatedPersonInfo>
        <relatedPersonInfo>
            <relatedPersonName>
                <firstName>Reza</firstName>
                <lastName>Dana</lastName>
            </relatedPersonName>
            <relatedPersonAddress>
                <street1>266 Main St., BLDG 2, STE 11</street1>
                <city>Medfield</city>
                <stateOrCountry>MA</stateOrCountry>
                <stateOrCountryDescription>MASSACHUSETTS</stateOrCountryDescription>
                <zipCode>02052</zipCode>
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            <relationshipClarification></relationshipClarification>
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        <relatedPersonInfo>
            <relatedPersonName>
                <firstName>Jiagang</firstName>
                <lastName>Zhao</lastName>
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            <relatedPersonAddress>
                <street1>266 Main St., BLDG 2, STE 11</street1>
                <city>Medfield</city>
                <stateOrCountry>MA</stateOrCountry>
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                <zipCode>02052</zipCode>
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        <relatedPersonInfo>
            <relatedPersonName>
                <firstName>David</firstName>
                <lastName>Neafus</lastName>
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            <relatedPersonAddress>
                <street1>266 Main St., BLDG 2, STE 11</street1>
                <city>Medfield</city>
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                <zipCode>02052</zipCode>
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            <relatedPersonRelationshipList>
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            <industryGroupType>Biotechnology</industryGroupType>
        </industryGroup>
        <issuerSize>
            <revenueRange>Decline to Disclose</revenueRange>
        </issuerSize>
        <federalExemptionsExclusions>
            <item>06b</item>
        </federalExemptionsExclusions>
        <typeOfFiling>
            <newOrAmendment>
                <isAmendment>true</isAmendment>
                <previousAccessionNumber>0001932386-23-000001</previousAccessionNumber>
            </newOrAmendment>
            <dateOfFirstSale>
                <value>2022-05-25</value>
            </dateOfFirstSale>
        </typeOfFiling>
        <durationOfOffering>
            <moreThanOneYear>true</moreThanOneYear>
        </durationOfOffering>
        <typesOfSecuritiesOffered>
            <isSecurityToBeAcquiredType>true</isSecurityToBeAcquiredType>
            <isOtherType>true</isOtherType>
            <descriptionOfOtherType>Simple Agreement for Future Equity</descriptionOfOtherType>
        </typesOfSecuritiesOffered>
        <businessCombinationTransaction>
            <isBusinessCombinationTransaction>true</isBusinessCombinationTransaction>
            <clarificationOfResponse></clarificationOfResponse>
        </businessCombinationTransaction>
        <minimumInvestmentAccepted>0</minimumInvestmentAccepted>
        <salesCompensationList></salesCompensationList>
        <offeringSalesAmounts>
            <totalOfferingAmount>3500000</totalOfferingAmount>
            <totalAmountSold>3065000</totalAmountSold>
            <totalRemaining>435000</totalRemaining>
            <clarificationOfResponse></clarificationOfResponse>
        </offeringSalesAmounts>
        <investors>
            <hasNonAccreditedInvestors>false</hasNonAccreditedInvestors>
            <totalNumberAlreadyInvested>21</totalNumberAlreadyInvested>
        </investors>
        <salesCommissionsFindersFees>
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                <dollarAmount>0</dollarAmount>
            </salesCommissions>
            <findersFees>
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            </findersFees>
            <clarificationOfResponse></clarificationOfResponse>
        </salesCommissionsFindersFees>
        <useOfProceeds>
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                <dollarAmount>300000</dollarAmount>
                <isEstimate>true</isEstimate>
            </grossProceedsUsed>
            <clarificationOfResponse></clarificationOfResponse>
        </useOfProceeds>
        <signatureBlock>
            <authorizedRepresentative>false</authorizedRepresentative>
            <signature>
                <issuerName>SightStream Biotherapeutics, Inc.</issuerName>
                <signatureName>/s/ Jason Werner</signatureName>
                <nameOfSigner>Jason Werner</nameOfSigner>
                <signatureTitle>President</signatureTitle>
                <signatureDate>2023-06-12</signatureDate>
            </signature>
        </signatureBlock>
    </offeringData>
</edgarSubmission>
