Genome-Scale CRISPR Screens Identify Human Pluripotency-Specific Genes.

Autor:	Ihry RJ; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA. Electronic address: robert.ihry@novartis.com., Salick MR; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Insitro, South San Francisco, CA 94080, USA., Ho DJ; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Sondey M; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Abbvie, Cambridge, MA 02139, USA., Kommineni S; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Casma Therapeutics, Cambridge, MA 02139, USA., Paula S; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Raymond J; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Henry B; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Frias E; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Wang Q; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Worringer KA; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Ye C; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Blueprint Medicines, Cambridge, MA 02139, USA., Russ C; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Reece-Hoyes JS; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Altshuler RC; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Randhawa R; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Axcella Health, Cambridge, MA 02139, USA., Yang Z; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; University of Massachusetts Medical School, Worcester, MA 01655, USA., McAllister G; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Sana Biotechnology, Cambridge, MA 02139, USA., Hoffman GR; Department of Chemical Biology and Therapeutics, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Sana Biotechnology, Cambridge, MA 02139, USA., Dolmetsch R; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA., Kaykas A; Department of Neuroscience, Novartis Institutes for Biomedical Research, Cambridge, MA 02139, USA; Insitro, South San Francisco, CA 94080, USA. Electronic address: akaykas@gmail.com.
Jazyk:	angličtina
Zdroj:	Cell reports [Cell Rep] 2019 Apr 09; Vol. 27 (2), pp. 616-630.e6.
DOI:	10.1016/j.celrep.2019.03.043
Abstrakt:	Human pluripotent stem cells (hPSCs) generate a variety of disease-relevant cells that can be used to improve the translation of preclinical research. Despite the potential of hPSCs, their use for genetic screening has been limited by technical challenges. We developed a scalable and renewable Cas9 and sgRNA-hPSC library in which loss-of-function mutations can be induced at will. Our inducible mutant hPSC library can be used for multiple genome-wide CRISPR screens in a variety of hPSC-induced cell types. As proof of concept, we performed three screens for regulators of properties fundamental to hPSCs: their ability to self-renew and/or survive (fitness), their inability to survive as single-cell clones, and their capacity to differentiate. We identified the majority of known genes and pathways involved in these processes, as well as a plethora of genes with unidentified roles. This resource will increase the understanding of human development and genetics. This approach will be a powerful tool to identify disease-modifying genes and pathways. (Copyright © 2019 The Author(s). Published by Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
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