Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera
Turner, George F.
Smith, Alan M.
Miska, Eric A.
Juntti, Scott A.
Royal Society Open Science
The Royal Society
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Clark, B., Elkin, J., Marconi, A., Turner, G. F., Smith, A. M., Joyce, D., Miska, E. A., et al. (2022). Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera. Royal Society Open Science, 9 (4) https://doi.org/10.1098/rsos.220077
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 cichlids radiation, establishing genome editing in this species will facilitate investigating speciation, adaptation and trait diversification in this textbook radiation.
Organismal and evolutionary biology, Research articles, cichlids, CRISPR/Cas9, adaptive radiation, functional genetics, evo-devo
Cancer Research UK (C13474/A18583, C6946/A14492)
Human Frontier Science Program (RGY0079/2018)
Natural Environment Research Council (NE/R01504X/1)
Wellcome Trust (092096/Z/10/Z, 102175/Z/13/Z, 219475/Z/19/Z, 222279/Z/20/Z)
External DOI: https://doi.org/10.1098/rsos.220077
This record's URL: https://www.repository.cam.ac.uk/handle/1810/337076