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

Published version
Peer-reviewed

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Type

Article

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Authors

Clark, Bethan 
Elkin, Joel 
Marconi, Aleksandra 
Turner, George F. 
Smith, Alan M. 

Abstract

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.

Description

Keywords

Organismal and evolutionary biology, Research articles, cichlids, CRISPR/Cas9, adaptive radiation, functional genetics, evo-devo

Journal Title

Royal Society Open Science

Conference Name

Journal ISSN

2054-5703

Volume Title

9

Publisher

The Royal Society
Sponsorship
Cancer Research UK (C13474/A18583, C6946/A14492)
Human Frontier Science Program (RGY0079/2018)
NSF (IOS-1825723)
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)