清脆的
Cas9
生物
基因组编辑
基因
遗传学
突变
计算生物学
等位基因
基因靶向
错义突变
功能(生物学)
突变
作者
Annelise Cassidy,Emin Kuliyev,Destinée B. Thomas,Hanying Chen,S. William Pelletier
出处
期刊:Methods in Enzymology
日期:2022-01-01
卷期号:: 775-812
被引量:5
标识
DOI:10.1016/bs.mie.2022.03.053
摘要
Allelic series are extremely valuable genetic tools to study gene function and identify essential structural features of gene products. In mice, allelic series have been engineered using conventional gene targeting in embryonic stem cells or chemical mutagenesis. While these approaches have provided valuable information about the function of genes, they remain cumbersome. Modern approaches such as CRISPR-Cas9 technologies now allow for the precise and cost-effective generation of mouse models with specific mutations, facilitating the development of allelic series. Here, we describe procedures for the generation of three types of mutations used to dissect protein function in vivo using CRISPR-Cas9 technology. This step-by-step protocol describes the generation of missense mutations, large in-frame deletions, and insertions of genetic material using SCY1-like 1 (Scyl1) as a model gene.
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