清脆的
功能基因组学
Cas9
基因敲除
生物
计算生物学
基因
基因组编辑
引导RNA
遗传筛选
基因组学
药物发现
功能(生物学)
遗传学
表型
基因组
生物信息学
作者
Michael Aregger,Megha Chandrashekhar,Amy H.Y. Tong,Katherine Chan,Jason Moffat
出处
期刊:Methods in molecular biology
日期:2018-10-16
卷期号:: 169-188
被引量:25
标识
DOI:10.1007/978-1-4939-8805-1_15
摘要
CRISPR-Cas9 technology provides a simple way to introduce targeted mutations into mammalian cells to induce loss-of-function phenotypes. The CRISPR-Cas9 system has now successfully been applied for genetic screens in many cell types, providing a powerful tool for functional genomics with manifold applications. Genome-wide guide-RNA (gRNA) libraries allow facile generation of a pool of cells, each harboring a gene knockout mutation that can be used for the study of gene function, pathway analysis or the identification of genes required for cellular fitness. Furthermore, CRISPR genetic screens can be applied for the discovery of genes whose knockout sensitizes cells to drug treatments or mediates drug resistance. Here, we provide a detailed protocol discussing the necessary steps for the successful performance of pooled CRISPR-Cas9 screens.
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