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
Related Resources
PubMed ID
30970262
Repository Citation
Ihry, Robert J.; Yang, Zinger; and Kaykas, Ajamete, "Genome-Scale CRISPR Screens Identify Human Pluripotency-Specific Genes" (2019). Open Access Articles. 3821.
https://escholarship.umassmed.edu/oapubs/3821
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.
Included in
Cell Biology Commons, Cells Commons, Computational Biology Commons, Developmental Biology Commons, Genetics Commons, Genomics Commons, Investigative Techniques Commons, Molecular Genetics Commons, Nucleic Acids, Nucleotides, and Nucleosides Commons
Comments
Full author list omitted for brevity. For the full list of authors, see article.