Engineering Forward Genetics into Cultured Cancer Cells for Chemical Target Identification.

Autor:	Povedano JM; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., Liou J; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., Wei D; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., Srivatsav A; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., Kim J; Department of Population and Data Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., Xie Y; Department of Population and Data Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., Nijhawan D; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA., McFadden DG; Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Harold C. Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA. Electronic address: david.mcfadden@utsouthwestern.edu.
Jazyk:	angličtina
Zdroj:	Cell chemical biology [Cell Chem Biol] 2019 Sep 19; Vol. 26 (9), pp. 1315-1321.e3. Date of Electronic Publication: 2019 Jul 11.
DOI:	10.1016/j.chembiol.2019.06.006
Abstrakt:	Target identification for biologically active small molecules remains a major barrier for drug discovery. Cancer cells exhibiting defective DNA mismatch repair (dMMR) have been used as a forward genetics system to uncover compound targets. However, this approach has been limited by the dearth of cancer cell lines that harbor naturally arising dMMR. Here, we establish a platform for forward genetic screening using CRISPR/Cas9 to engineer dMMR into mammalian cells. We demonstrate the utility of this approach to identify mechanisms of drug action in mouse and human cancer cell lines using in vitro selections against three cellular toxins. In each screen, compound-resistant alleles emerged in drug-resistant clones, supporting the notion that engineered dMMR enables forward genetic screening in mammalian cells. (Copyright © 2019 Elsevier Ltd. All rights reserved.)
Databáze:	MEDLINE
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