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Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera

cam.issuedOnline2022-04-20
dc.contributor.authorClark, Bethan
dc.contributor.authorElkin, Joel
dc.contributor.authorMarconi, Aleksandra
dc.contributor.authorTurner, George F
dc.contributor.authorSmith, Alan M
dc.contributor.authorJoyce, Domino
dc.contributor.authorMiska, Eric A
dc.contributor.authorJuntti, Scott A
dc.contributor.authorSantos, M Emília
dc.contributor.orcidJoyce, Domino [0000-0002-4253-5623]
dc.contributor.orcidSantos, M Emília [0000-0003-3158-7935]
dc.date.accessioned2022-04-20T09:00:15Z
dc.date.available2022-04-20T09:00:15Z
dc.date.issued2022-04
dc.date.submitted2022-01-20
dc.date.updated2022-04-20T09:00:14Z
dc.description.abstract<jats:p> Identifying genetic loci underlying trait variation provides insights into the mechanisms of diversification, but demonstrating causality and characterizing the role of genetic loci requires testing candidate gene function, often in non-model species. Here we establish CRISPR/Cas9 editing in <jats:italic>Astatotilapia calliptera</jats:italic> , a generalist cichlid of the remarkably diverse Lake Malawi radiation. By targeting the gene <jats:italic>oca2</jats:italic> required for melanin synthesis in other vertebrate species, we show efficient editing and germline transmission. Gene edits include indels in the coding region, probably a result of non-homologous end joining, and a large deletion in the 3′ untranslated region due to homology-directed repair. We find that <jats:italic>oca2</jats:italic> knock-out <jats:italic>A. calliptera</jats:italic> lack melanin, which may be useful for developmental imaging in embryos and studying colour pattern formation in adults. As <jats:italic>A. calliptera</jats:italic> resembles the presumed generalist ancestor of the Lake Malawi cichlids radiation, establishing genome editing in this species will facilitate investigating speciation, adaptation and trait diversification in this textbook radiation. </jats:p>
dc.identifier.doi10.17863/CAM.83672
dc.identifier.eissn2054-5703
dc.identifier.issn2054-5703
dc.identifier.otherrsos220077
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/336253
dc.languageen
dc.publisherThe Royal Society
dc.subjectOrganismal and evolutionary biology
dc.subjectResearch articles
dc.subjectcichlids
dc.subjectCRISPR/Cas9
dc.subjectadaptive radiation
dc.subjectfunctional genetics
dc.subjectevo-devo
dc.title<i>Oca2</i> targeting using CRISPR/Cas9 in the Malawi cichlid <i>Astatotilapia calliptera</i>
dc.typeArticle
dcterms.dateAccepted2022-03-22
prism.issueIdentifier4
prism.publicationNameRoyal Society Open Science
prism.volume9
pubs.funder-project-idCancer Research UK (C13474/A18583, C6946/A14492)
pubs.funder-project-idHuman Frontier Science Program (RGY0079/2018)
pubs.funder-project-idNSF (IOS-1825723)
pubs.funder-project-idNatural Environment Research Council (NE/R01504X/1)
pubs.funder-project-idWellcome Trust (092096/Z/10/Z, 102175/Z/13/Z, 219475/Z/19/Z, 222279/Z/20/Z)
rioxxterms.licenseref.urihttp://creativecommons.org/licenses/by/4.0/
rioxxterms.versionVoR
rioxxterms.versionofrecord10.1098/rsos.220077

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