Rapid and tunable method to temporally control gene editing based on conditional Cas9 stabilization Academic Article uri icon

Overview

MeSH Major

  • Alternative Splicing
  • Angiogenesis Inhibitors
  • Exons
  • RNA Splice Sites
  • RNA-Binding Proteins
  • Vascular Endothelial Growth Factor A

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.

publication date

  • February 22, 2017

Research

keywords

  • Academic Article

Identity

Language

  • eng

PubMed Central ID

  • PMC5322564

Digital Object Identifier (DOI)

  • 10.1038/ncomms14370

PubMed ID

  • 28224990

Additional Document Info

start page

  • 14370

volume

  • 8