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


Cancer Medicine & Anti Cancer Drugs

Author(s): Smurnyy Y, Cai M, Wu H, McWhinnie E, Tallarico JA,

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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. This article was published in Nat Chem Biol and referenced in Cancer Medicine & Anti Cancer Drugs

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