title: 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.
authors.affiliation: Addgene, Cambridge, Massachusetts.
authors.forename: Joel R
authors.initials: JR
authors.lastname: McDade
authors.affiliation: Addgene, Cambridge, Massachusetts.
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Waxmonsky
authors.affiliation: Addgene, Cambridge, Massachusetts.
authors.forename: Lianna E
authors.initials: LE
authors.lastname: Swanson
authors.affiliation: Addgene, Cambridge, Massachusetts.
authors.forename: Melina
authors.initials: M
authors.lastname: Fan
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D015723:Gene Library; D018390:Gene Targeting / Q000379:methods; D005818:Genetic Engineering / Q000379:methods; D016678:Genome; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009154:Mutation; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems
keywords: CRISPR/Cas9; genome-wide screening; pooled libraries