Shalem Ophir, Sanjana Neville E, Zhang Feng
Broad Institute of MIT and Harvard, 7 Cambridge Center, Cambridge, Massachusetts 02142, USA; McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, and Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
Nat Rev Genet. 2015 May;16(5):299-311. doi: 10.1038/nrg3899. Epub 2015 Apr 9.
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.
正向遗传学筛选是发现遗传元件并对其进行功能注释的强大工具。最近，RNA引导的CRISPR（规律成簇的间隔短回文重复序列）相关的Cas9核酸酶已与基因组规模的向导RNA文库相结合，用于无偏差的表型筛选。在本综述中，我们描述了使用Cas9进行基因组规模筛选的最新进展，包括使基因组位点失活的敲除方法和调节转录活性的策略。我们讨论了筛选设计的实际问题，与RNA干扰（RNAi）筛选进行了比较，并概述了未来的应用和挑战。
