Rapid and Tunable Method to Temporally Control Gene Editing Based on Conditional Cas9 Stabilization

Overview

Journal Nat Commun

Specialty Biology

Date 2017 Feb 23

PMID 28224990

Citations 80

Authors

Serif Senturk

Nitin H Shirole

Dawid G Nowak

Vincenzo Corbo

Debjani Pal

Alexander Vaughan

David A Tuveson

Lloyd C Trotman

Justin B Kinney

Raffaella Sordella

Affiliations

Soon will be listed here.

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-ER, 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.

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