High-throughput functional genomics using CRISPR-Cas9.
High-throughput functional genomics using CRISPR-Cas9.
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DOI：
10.1038/nrg3899
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发表时间：
2015-05
期刊：
影响因子：
--
通讯作者：
Zhang F
中科院分区：
文献类型：
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作者：
Shalem O;Sanjana NE;Zhang F
正向遗传筛选是发现和功能注释遗传元件的有力工具。最近，RNA引导的CRISPR（成簇的规则间隔短回文重复序列）相关的Cas9核酸酶已与基因组规模的引导RNA文库组合用于无偏的表型筛选。在这篇综述中，我们描述了使用Cas9进行基因组规模筛选的最新进展，包括敲除基因组位点的方法和调节转录活性的策略。我们讨论了屏幕设计的实际方面，提供与RNA干扰（RNAi）筛选的比较，并概述了未来的应用和挑战。
Forward genetic screens are powerful tools for the discovery and functional annotation of genetic elements. Recently, the RNA-guided CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease has been combined with genome-scale guide RNA libraries for unbiased, phenotypic screening. In this Review, we describe recent advances using Cas9 for genome-scale screens, including knockout approaches that inactivate genomic loci and strategies that modulate transcriptional activity. We discuss practical aspects of screen design, provide comparisons with RNA interference (RNAi) screening, and outline future applications and challenges.