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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Cell Line / drug effects
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Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
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DNA Helicases / genetics
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DNA-Binding Proteins / genetics
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Diterpenes / pharmacology
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Drug Resistance / drug effects
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Drug Resistance / genetics
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Endonucleases / genetics*
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Epoxy Compounds / pharmacology
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HCT116 Cells
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High-Throughput Nucleotide Sequencing / methods*
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Humans
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Hypoxanthine Phosphoribosyltransferase / genetics
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Mammals
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Phenanthrenes / pharmacology
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Point Mutation
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Reproducibility of Results
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Thioguanine / pharmacology
Substances
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DNA-Binding Proteins
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Diterpenes
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Epoxy Compounds
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Phenanthrenes
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XPBC-ERCC-3 protein
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triptolide
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Hypoxanthine Phosphoribosyltransferase
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Endonucleases
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DNA Helicases
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Thioguanine