High-throughput functional genomics using CRISPR–Cas9

O Shalem, NE Sanjana, F Zhang - Nature Reviews Genetics, 2015 - nature.com
Nature Reviews Genetics, 2015nature.com
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 …
Abstract
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.
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