Practical Considerations for Using Pooled Lentiviral CRISPR Libraries
- PMID: 27366891
- DOI: 10.1002/cpmb.8
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.
Keywords: CRISPR/Cas9; genome-wide screening; pooled libraries.
Copyright © 2016 John Wiley & Sons, Inc.
References
Literature Cited
-
- 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.
-
- 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.
-
- 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.
-
- 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.
-
- 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.
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