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

    <schemaVersion>X0407</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2023-03-31</periodOfReport>

    <notSubjectToSection16>0</notSubjectToSection16>

    <issuer>
        <issuerCik>0001807166</issuerCik>
        <issuerName>Amesite Inc.</issuerName>
        <issuerTradingSymbol>AMST</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001199190</rptOwnerCik>
            <rptOwnerName>LOSH J MICHAEL</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>C/O AMESITE INC.</rptOwnerStreet1>
            <rptOwnerStreet2>607 SHELBY STREET, SUITE 700 PMB 214</rptOwnerStreet2>
            <rptOwnerCity>DETROIT</rptOwnerCity>
            <rptOwnerState>MI</rptOwnerState>
            <rptOwnerZipCode>48226</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>1</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>0</isTenPercentOwner>
            <isOther>0</isOther>
            <officerTitle></officerTitle>
            <otherText></otherText>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <derivativeTable>
        <derivativeTransaction>
            <securityTitle>
                <value>Restricted Stock Units</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <footnoteId id="F1"/>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2023-03-31</value>
            </transactionDate>
            <deemedExecutionDate></deemedExecutionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionTimeliness></transactionTimeliness>
            <transactionAmounts>
                <transactionShares>
                    <value>5556</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>0</value>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F2"/>
            </exerciseDate>
            <expirationDate>
                <footnoteId id="F2"/>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>5556</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>22222</value>
                    <footnoteId id="F3"/>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>I</value>
                </directOrIndirectOwnership>
                <natureOfOwnership>
                    <value>J. Michael Losh Irrevocable Qualified Annuity Trust #7</value>
                </natureOfOwnership>
            </ownershipNature>
        </derivativeTransaction>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">Each restricted stock unit (&quot;RSU&quot;) represents a contingent right to receive one share of AMST common stock and/or the cash equivalent of such share.</footnote>
        <footnote id="F2">The reported securities were issued as &quot;deferred stock units&quot; in lieu of cash compensation earned for services as a director, and the number of deferred stock units received was calculated based on the closing share price on the last day of the quarter. The deferred stock units or cash equivalent will be issued to the reporting person upon completion of service as a member of the board of directors or, if earlier, a change in control.</footnote>
        <footnote id="F3">On February 21, 2023, a 1-for-12 reverse stock split of the common stock of AMST was effected, resulting in the reporting person's ownership of one-twelfth (1/12) of the number of RSUs owned prior to such reverse stock split.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Sherrie Farrell, Attorney-in-Fact</signatureName>
        <signatureDate>2023-04-04</signatureDate>
    </ownerSignature>
</ownershipDocument>
