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

Serif Senturk, Nitin H. Shirole, Dawid G. Nowak, Vincenzo Corbo, Debjani Pal, Alexander Vaughan, David A. Tuveson, Lloyd C. Trotman, Justin B. Kinney and Raffaella Sordella ()
Additional contact information
Serif Senturk: Cold Spring Harbor Laboratory
Nitin H. Shirole: Cold Spring Harbor Laboratory
Dawid G. Nowak: Cold Spring Harbor Laboratory
Vincenzo Corbo: Cold Spring Harbor Laboratory
Debjani Pal: Cold Spring Harbor Laboratory
Alexander Vaughan: Cold Spring Harbor Laboratory
David A. Tuveson: Cold Spring Harbor Laboratory
Lloyd C. Trotman: Cold Spring Harbor Laboratory
Justin B. Kinney: Cold Spring Harbor Laboratory
Raffaella Sordella: Cold Spring Harbor Laboratory

Nature Communications, 2017, vol. 8, issue 1, 1-10

Abstract: 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.

Date: 2017
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DOI: 10.1038/ncomms14370

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