Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2016 Jul 1;115:31.5.1-31.5.13.
doi: 10.1002/cpmb.8.

Practical Considerations for Using Pooled Lentiviral CRISPR Libraries

Affiliations
Review

Practical Considerations for Using Pooled Lentiviral CRISPR Libraries

Joel R McDade et al. Curr Protoc Mol Biol. .

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.

Keywords: CRISPR/Cas9; genome-wide screening; pooled libraries.

Similar articles

See all similar articles

Cited by 1 article

  • Functional Genomics via CRISPR-Cas.
    Ford K, McDonald D, Mali P. Ford K, et al. J Mol Biol. 2019 Jan 4;431(1):48-65. doi: 10.1016/j.jmb.2018.06.034. Epub 2018 Jun 28. J Mol Biol. 2019. PMID: 29959923 Free PMC article. Review.

References

Literature Cited

    1. Anders, C. , Niewoehner, O. , Duerst, A. , and Jinek, M. 2014. Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature 513:569-573. doi: 10.1038/nature13579.
    1. Bassett, A.R. , Kong, L. , and Liu, J.L. 2015. A genome-wide CRISPR library for high-throughput genetic screening in Drosophila cells. J. Genet. Genomics 42:301-309. doi: 10.1016/j.jgg.2015.03.011.
    1. Boettcher, M. and McManus, M.T. 2015. Choosing the right tool for the job: RNAi, TALEN, or CRISPR. Cell 4:575-585. doi: 10.1016/j.molcel.2015.04.028.
    1. Chavez, A. , Scheiman, J. , Vora, S. , Pruitt, B.W. , Tuttle, M. , Iyer, E.P.R. , Lin, S. , Kiani, S. , Guzman, C.D. , Wiegand, D.J. , Ter-Ovanesyan, D. , Braff, J.L. , Davidsohn, N. , Housden, B.E. , Perrimon, N. , Weiss, R. , Aach, J. , Collins, J.J. , and Church, G.M. 2015. Highly efficient Cas9-mediated transcriptional programming. Nat. Methods 12:326-328. doi: 10.1038/nmeth.3312.
    1. Chen, S. , Sanjana, N.E. , Zheng, K. , Shalem, O. , Lee, K. , Shi, X. , Scott, D.A. , Song, J. , Pan, J.Q. , Weissleder, R. , Lee, H. , Zhang, F. , and Sharp, P.A. 2015. Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis. Cell 160:1246-1260. doi: 10.1016/j.cell.2015.02.038.

LinkOut - more resources

Feedback