Rapid Generation of Pigment Free, Immobile Zebrafish Embryos and Larvae in Any Genetic Background Using CRISPR-Cas9 dgRNPs.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2021-08-01 Epub Date: 2021-06-02 DOI:10.1089/zeb.2021.0011

Andrew E Davis, Daniel Castranova, Brant M Weinstein

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引用次数: 4

Abstract

The ability to carry out high-resolution, high-magnification optical imaging of living animals is one of the most attractive features of the zebrafish as a model organism. However, increasing amounts of pigmentation as development proceeds and difficulties in maintaining sustained immobilization of healthy, living animals remain challenges for live imaging. Chemical treatments can be used to suppress pigment formation and movement, but these treatments can lead to developmental defects. Genetic mutants can also be used to eliminate pigment formation and immobilize animals, but maintaining these mutants in lines carrying other combinations of transgenes and mutants is difficult and laborious. In this study, we show that CRISPR duplex guide ribonucleoproteins (dgRNPs) targeting the slc45a2 (albino) and chrna1 (nic1) genes can be used to efficiently suppress pigment formation in and immobilize F0 injected animals. CRISPR dgRNPs can be used to generate pigment-free, immobile zebrafish embryos and larvae in any transgenic and/or mutant-carrying background, greatly facilitating high-resolution imaging and analysis of the many transgenic and mutant lines available in the zebrafish.

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利用CRISPR-Cas9 dgRNPs快速生成任何遗传背景下无色素、不动的斑马鱼胚胎和幼虫

对活体动物进行高分辨率、高倍率光学成像的能力是斑马鱼作为模式生物最吸引人的特征之一。然而，随着发育的进行，色素沉着量的增加以及维持健康活体动物持续固定的困难仍然是活体成像的挑战。化学处理可以抑制色素的形成和运动，但这些处理可能导致发育缺陷。基因突变体也可用于消除色素形成和固定动物，但在携带其他转基因和突变体组合的品种中维持这些突变体是困难和费力的。在本研究中，我们发现靶向slc45a2(白化病)和chrna1 (nic1)基因的CRISPR双导核糖核蛋白(dgRNPs)可有效抑制F0注射动物的色素形成并使其固定化。CRISPR dgRNPs可用于在任何转基因和/或携带突变体的背景下产生无色素、不动的斑马鱼胚胎和幼虫，极大地促进了对斑马鱼中许多转基因和突变系的高分辨率成像和分析。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.