Practical Considerations for Using Pooled Lentiviral CRISPR Libraries

Abstract CRISPR/Cas9 technology is ideally suited for genome‐wide screening applications due to the ease of generating guide RNAs (gRNAs) and the versatility of Cas9 or Cas9 derivatives to knockout, repress, or activate expression of target genes. Several pooled lentiviral CRISPR libraries have been developed and are now publicly available, but while using CRISPR/Cas9 for genetic experiments has become widely adopted, genome‐wide screening experiments remain technically challenging. This review covers the basics of CRISPR/Cas9, describes several publicly available CRISPR libraries, and provides a general protocol for conducting genome‐wide screening experiments using CRISPR/Cas9. © 2016 by John Wiley & Sons, Inc.