Details

Autor(en) / Beteiligte

• Senturk, Serif
• Shirole, Nitin H
• Nowak, Dawid G
• Corbo, Vincenzo
• Pal, Debjani
• Vaughan, Alexander
• Tuveson, David A
• Trotman, Lloyd C
• Kinney, Justin B
• Sordella, Raffaella

Titel

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

Ist Teil von

• Nature communications, 2017-02, Vol.8 (1), p.14370-14370, Article 14370

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2017

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.