High-throughput Functional Genomics Using CRISPR-Cas9

Nat Rev Genet. 2015 May;16(5):299-311. doi: 10.1038/nrg3899. Epub 2015 Apr 9.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • CRISPR-Cas Systems*
  • Gene Knockout Techniques
  • Gene Library
  • Genetic Testing / methods
  • Genomics / methods*
  • High-Throughput Nucleotide Sequencing / methods
  • Humans
  • Mice
  • Models, Genetic
  • RNA Interference