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

Clark, Bethan; Elkin, Joel; Marconi, Aleksandra; Turner, George F.; Smith, Alan M.; Joyce, Domino; Miska, Eric A.; Juntti, Scott A.; Santos, M. Emília

Authors

Bethan Clark

Joel Elkin

Aleksandra Marconi

George F. Turner

Alan M. Smith

Eric A. Miska

Scott A. Juntti

M. Emília Santos



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 cichlids radiation, establishing genome editing in this species will facilitate investigating speciation, adaptation and trait diversification in this textbook radiation.

Citation

Clark, B., Elkin, J., Marconi, A., Turner, G. F., Smith, A. M., Joyce, D., …Santos, M. E. (2022). Oca2 targeting using CRISPR/Cas9 in the Malawi cichlid Astatotilapia calliptera. Royal Society Open Science, 9(4), Article 220077. https://doi.org/10.1098/rsos.220077

Journal Article Type Article
Acceptance Date Mar 22, 2022
Online Publication Date Apr 20, 2022
Publication Date 2022-04
Deposit Date May 31, 2022
Publicly Available Date Oct 27, 2022
Journal Royal Society Open Science
Electronic ISSN 2054-5703
Publisher The Royal Society
Peer Reviewed Peer Reviewed
Volume 9
Issue 4
Article Number 220077
DOI https://doi.org/10.1098/rsos.220077
Keywords Evo-devo; CRISPR/Cas9; Functional genetics; Adaptive radiation; Cichlids
Public URL https://hull-repository.worktribe.com/output/4000096

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http://creativecommons.org/licenses/by/4.0

Copyright Statement
© 2022 The Authors.

Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.



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