Smurnyy, Yegor, Cai, Mi, Wu, Hua, Mc Whinnie, Elizabeth, Tallarico, John, Yang, Yi and Feng, Yan (2014) CRISPR Mediated Target Validation of ERCC3 as the Target of Triptolide. Nature Chemical Biology.

Abstract

Exome sequencing of Triptolide-resistant haploid cells identified mutations in TFIIH complex, with a majority of them in the ERCC3 subunit. These mutations turned out to be recessive, therefore difficult to validate using conventional cDNA overexpression approach. Here we adopted a novel CRISPR-based knock-in strategy which makes it very straightforward to re-introduce the recessive point mutations that confer drug resistance phenotype in cells. Coupled with mutagenesis and next generation sequencing, this novel gene-editing strategy can expedite small molecule target identification and validation in mammalian cells.

Item Type:	Article
Keywords:	CRISPR, Triptolide, ERCC3, XPB, Next generation sequencing, mutagenesis, drug-resistant, exome sequencing, CRISPR-mediated knock-in
Date Deposited:	13 Oct 2015 13:13
Last Modified:	13 Oct 2015 13:13
URI:	https://oak.novartis.com/id/eprint/20529