RT Journal Article SR Electronic T1 A rapid and tunable method to temporally control Cas9 expression enables the identification of essential genes and the interrogation of functional gene interactions in vitro and in vivo JF bioRxiv FD Cold Spring Harbor Laboratory SP 023366 DO 10.1101/023366 A1 Serif Senturk A1 Nitin H. Shirole A1 Dawid D. Nowak A1 Vincenzo Corbo A1 Alexander Vaughan A1 David A. Tuveson A1 Lloyd C. Trotman A1 Adam Kepecs A1 Frank Stegmeier A1 Raffaella Sordella YR 2015 UL http://biorxiv.org/content/early/2015/07/28/023366.abstract AB The Cas9/CRISPR system is a powerful tool for studying gene function. Here we describe a method that allows temporal control of Cas9/CRISPER activity based on conditional Cas9 destabilization. We demonstrate that fusing an FKBP12-derived destabilizing domain to Cas9 (DD-CAS9) enables conditional Cas9 expression in vitro in the presence of an FKBP12 synthetic ligand and temporal control of gene-editing. Further, we show that this strategy can be easily adapted to co-express, from the same promoter, DD-Cas9 with any other gene of interest, without the latter being co-modulated. 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 identification of essential genes and the interrogation of genes functional interactions.