秀丽隐杆线虫中CRISPR-Cas9引导的基因组工程
CRISPR-Cas9-Guided Genome Engineering in C. elegans.
作者信息
Kim Hyun-Min, Colaiácovo Monica P
机构信息
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
出版信息
Curr Protoc Mol Biol. 2016 Jul 1;115:31.7.1-31.7.18. doi: 10.1002/cpmb.7.
Abstract
The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system is successfully being used for efficient and targeted genome editing in various organisms, including the nematode C. elegans. Recent studies have developed various CRISPR-Cas9 approaches to enhance genome engineering via two major DNA double-strand break repair pathways: non-homologous end joining and homologous recombination. Here we describe a protocol for Cas9-mediated C. elegans genome editing together with single guide RNA (sgRNA) and repair template cloning, as well as injection methods required for delivering Cas9, sgRNAs, and repair template DNA into the C. elegans germline. © 2016 by John Wiley & Sons, Inc.
摘要
CRISPR（规律成簇间隔短回文重复序列）-Cas（CRISPR相关）系统已成功用于多种生物体的高效靶向基因组编辑，包括线虫秀丽隐杆线虫。最近的研究开发了各种CRISPR-Cas9方法，通过两种主要的DNA双链断裂修复途径来增强基因组工程：非同源末端连接和同源重组。本文我们描述了一种用于Cas9介导的秀丽隐杆线虫基因组编辑的方案，以及单向导RNA（sgRNA）和修复模板克隆，还有将Cas9、sgRNAs和修复模板DNA导入秀丽隐杆线虫种系所需的注射方法。© 2016约翰威立父子公司版权所有
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