Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization

Senturk, S., Shirole, N. H., Nowak, D. G., Corbo, V., Pal, D., Vaughan, A., Tuveson, D. A., Trotman, L. C., Kinney, J. B., Sordella, R. (February 2017) Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization. Nat Commun, 8. p. 14370. ISSN 2041-1723 (Public Dataset)

URL: https://www.ncbi.nlm.nih.gov/pubmed/28224990
DOI: 10.1038/ncomms14370

Abstract

The CRISPR/Cas9 system is a powerful tool for studying gene function. Here, we describe a method that allows temporal control of CRISPR/Cas9 activity based on conditional Cas9 destabilization. We demonstrate that fusing an FKBP12-derived destabilizing domain to Cas9 (DD-Cas9) enables conditional Cas9 expression and temporal control of gene editing in the presence of an FKBP12 synthetic ligand. This system can be easily adapted to co-express, from the same promoter, DD-Cas9 with any other gene of interest without co-modulation of the latter. In particular, when co-expressed with inducible Cre-ERT2, our system enables parallel, independent manipulation of alleles targeted by Cas9 and traditional recombinase with single-cell specificity. We anticipate this platform will be used for the systematic characterization and identification of essential genes, as well as the investigation of the interactions between functional genes.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics
Investigative techniques and equipment > CRISPR-Cas9
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression
CSHL Authors:
Communities: CSHL labs > Kinney lab
CSHL labs > Sordella lab
CSHL labs > Trotman lab
CSHL labs > Tuveson lab
CSHL Cancer Center Program > Signal Transduction
Depositing User: Matt Covey
Date: 22 February 2017
Date Deposited: 02 Mar 2017 17:51
Last Modified: 05 Mar 2018 19:55
PMCID: PMC5322564
Related URLs:
Dataset ID:
URI: http://repository.cshl.edu/id/eprint/34136

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