Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera
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 Astatot...
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| Format: | Article |
| Language: | English |
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The Royal Society
2022-04-01
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| Series: | Royal Society Open Science |
| Subjects: | |
| Online Access: | https://royalsocietypublishing.org/doi/10.1098/rsos.220077 |
| _version_ | 1797837548759810048 |
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| author | Bethan Clark Joel Elkin Aleksandra Marconi George F. Turner Alan M. Smith Domino Joyce Eric A. Miska Scott A. Juntti M. Emília Santos |
| author_facet | Bethan Clark Joel Elkin Aleksandra Marconi George F. Turner Alan M. Smith Domino Joyce Eric A. Miska Scott A. Juntti M. Emília Santos |
| author_sort | Bethan Clark |
| collection | DOAJ |
| description | 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 Astatotilapia calliptera, a generalist cichlid of the remarkably diverse Lake Malawi radiation. By targeting the gene oca2 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 oca2 knock-out A. calliptera lack melanin, which may be useful for developmental imaging in embryos and studying colour pattern formation in adults. As A. calliptera resembles the presumed generalist ancestor of the Lake Malawi cichlid radiation, establishing genome editing in this species will facilitate investigating speciation, adaptation and trait diversification in this textbook radiation. |
| first_indexed | 2024-04-09T15:27:35Z |
| format | Article |
| id | doaj.art-0c0edf7081cd464e9161fca75632c650 |
| institution | Directory Open Access Journal |
| issn | 2054-5703 |
| language | English |
| last_indexed | 2024-04-09T15:27:35Z |
| publishDate | 2022-04-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Royal Society Open Science |
| spelling | doaj.art-0c0edf7081cd464e9161fca75632c6502023-04-28T11:05:35ZengThe Royal SocietyRoyal Society Open Science2054-57032022-04-019410.1098/rsos.220077Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia callipteraBethan Clark0Joel Elkin1Aleksandra Marconi2George F. Turner3Alan M. Smith4Domino Joyce5Eric A. Miska6Scott A. Juntti7M. Emília Santos8Department of Zoology, University of Cambridge, UKDepartment of Zoology, University of Cambridge, UKDepartment of Zoology, University of Cambridge, UKSchool of Natural Sciences, Bangor University, Gwynedd LL57 2TH, UKDepartment of Biological and Marine Sciences, University of Hull, UKDepartment of Biological and Marine Sciences, University of Hull, UKDepartment of Genetics, University of Cambridge, UKUniversity of Maryland, USADepartment of Zoology, University of Cambridge, UKIdentifying 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 Astatotilapia calliptera, a generalist cichlid of the remarkably diverse Lake Malawi radiation. By targeting the gene oca2 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 oca2 knock-out A. calliptera lack melanin, which may be useful for developmental imaging in embryos and studying colour pattern formation in adults. As A. calliptera resembles the presumed generalist ancestor of the Lake Malawi cichlid radiation, establishing genome editing in this species will facilitate investigating speciation, adaptation and trait diversification in this textbook radiation.https://royalsocietypublishing.org/doi/10.1098/rsos.220077cichlidsCRISPR/Cas9adaptive radiationfunctional geneticsevo-devo |
| spellingShingle | Bethan Clark Joel Elkin Aleksandra Marconi George F. Turner Alan M. Smith Domino Joyce Eric A. Miska Scott A. Juntti M. Emília Santos Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera Royal Society Open Science cichlids CRISPR/Cas9 adaptive radiation functional genetics evo-devo |
| title | Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera |
| title_full | Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera |
| title_fullStr | Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera |
| title_full_unstemmed | Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera |
| title_short | Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera |
| title_sort | oca2 targeting using crispr cas9 in the malawi cichlid astatotilapia calliptera |
| topic | cichlids CRISPR/Cas9 adaptive radiation functional genetics evo-devo |
| url | https://royalsocietypublishing.org/doi/10.1098/rsos.220077 |
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