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CRISPR Mediated Target Validation of ERCC3 as the Target of Triptolide

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.


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


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