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Nat Chem Biol. 2014 Aug;10(8):623-5. doi: 10.1038/nchembio.1550. Epub 2014 Jun 15.

DNA sequencing and CRISPR-Cas9 gene editing for target validation in mammalian cells.

Author information

1
Developmental and Molecular Pathways, Novartis Institutes for Biomedical Research, Cambridge, Massachusetts, USA.

Abstract

Identification and validation of drug-resistant mutations can provide important insights into the mechanism of action of a compound. Here we demonstrate the feasibility of such an approach in mammalian cells using next-generation sequencing of drug-resistant clones and CRISPR-Cas9-mediated gene editing on two drug-target pairs, 6-thioguanine-HPRT1 and triptolide-ERCC3. We showed that disrupting functional HPRT1 allele or introducing ERCC3 point mutations by gene editing can confer drug resistance in cells.

PMID:
24929529
DOI:
10.1038/nchembio.1550
[Indexed for MEDLINE]

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