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在马拉维丽鱼中使用CRISPR/Cas9进行靶向操作。

targeting using CRISPR/Cas9 in the Malawi cichlid .

作者信息

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

机构信息

Department of Zoology, University of Cambridge, UK.

School of Natural Sciences, Bangor University, Gwynedd LL57 2TH, UK.

出版信息

R Soc Open Sci. 2022 Apr 20;9(4):220077. doi: 10.1098/rsos.220077. eCollection 2022 Apr.

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 , a generalist cichlid of the remarkably diverse Lake Malawi radiation. By targeting the gene 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 knock-out lack melanin, which may be useful for developmental imaging in embryos and studying colour pattern formation in adults. As 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.

摘要

识别性状变异背后的基因座有助于深入了解物种分化的机制,但要证明因果关系并描述基因座的作用,通常需要在非模式物种中测试候选基因的功能。在这里,我们在马拉维湖辐射中一种具有广泛适应性的丽鱼科鱼类中建立了CRISPR/Cas9编辑技术。通过靶向其他脊椎动物物种中黑色素合成所需的基因,我们展示了高效的编辑和种系传递。基因编辑包括编码区的插入缺失,这可能是非同源末端连接的结果,以及由于同源定向修复导致的3'非翻译区的大片段缺失。我们发现该基因敲除个体缺乏黑色素,这对于胚胎发育成像和研究成年个体的颜色模式形成可能有用。由于该物种类似于马拉维湖丽鱼科鱼类辐射的假定原始祖先,在该物种中建立基因组编辑将有助于研究这一典型辐射中的物种形成、适应和性状多样化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7fe3/9019512/9ca30848c727/rsos220077f01.jpg

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