UMMS Affiliation

Graduate School of Biomedical Sciences

Publication Date

2019-04-09

Document Type

Article

Disciplines

Cell Biology | Cells | Computational Biology | Developmental Biology | Genetics | Genomics | Investigative Techniques | Molecular Genetics | Nucleic Acids, Nucleotides, and Nucleosides

Abstract

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.

Keywords

CRISPR genome-wide screening, DDR, PAWR, PMAIP1, hESC, human pluripotent stem cells, iPSC

Rights and Permissions

© 2019 The Author(s). User License Creative Commons Attribution (CC BY 4.0).

DOI of Published Version

10.1016/j.celrep.2019.03.043

Source

Cell Rep. 2019 Apr 9;27(2):616-630.e6. doi: 10.1016/j.celrep.2019.03.043. Link to article on publisher's site

Journal/Book/Conference Title

Cell reports

Comments

Full author list omitted for brevity. For the full list of authors, see article.

Related Resources

Link to Article in PubMed

PubMed ID

30970262

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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