McDade Joel R, Waxmonsky Nicole C, Swanson Lianna E, Fan Melina
Addgene, Cambridge, Massachusetts.
Curr Protoc Mol Biol. 2016 Jul 1;115:31.5.1-31.5.13. doi: 10.1002/cpmb.8.
CRISPR/Cas9 technology is ideally suited for genome-wide screening applications due to the ease of generating guide RNAs (gRNAs) and the versatility of Cas9 or Cas9 derivatives to knockout, repress, or activate expression of target genes. Several pooled lentiviral CRISPR libraries have been developed and are now publicly available, but while using CRISPR/Cas9 for genetic experiments has become widely adopted, genome-wide screening experiments remain technically challenging. This review covers the basics of CRISPR/Cas9, describes several publicly available CRISPR libraries, and provides a general protocol for conducting genome-wide screening experiments using CRISPR/Cas9. © 2016 by John Wiley & Sons, Inc.
由于易于生成向导RNA（gRNA）以及Cas9或Cas9衍生物在敲除、抑制或激活靶基因表达方面的多功能性，CRISPR/Cas9技术非常适合全基因组筛选应用。已经开发了几个汇集的慢病毒CRISPR文库，现在可以公开获得，但是尽管使用CRISPR/Cas9进行基因实验已被广泛采用，但全基因组筛选实验在技术上仍然具有挑战性。本综述涵盖了CRISPR/Cas9的基础知识，描述了几个公开可用的CRISPR文库，并提供了使用CRISPR/Cas9进行全基因组筛选实验的一般方案。© 2016约翰威立父子公司版权所有。
