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Updated: Jan 26, 2026
10:24
Transfecting and Nucleofecting Human Induced Pluripotent Stem Cells
Published on: October 5, 2011
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Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies
Barbara Mair1, Jelena Tomic1, Sanna N Masud2
1Donnelly Centre, University of Toronto, Toronto, ON, Canada.
Cell Reports
|April 11, 2019
Summary
Researchers identified essential genes for human pluripotent stem cell (hPSC) fitness using CRISPR screens. These findings highlight context-dependent gene requirements and identify novel regulators of hPSC biology.
Area of Science:
- Stem Cell Biology
- Genetics
- Genomics
Background:
- Human pluripotent stem cells (hPSCs) are crucial for disease modeling and regenerative medicine.
- Understanding pluripotency and differentiation requires identifying genes essential for hPSC fitness (cell reproduction).
Purpose of the Study:
- To systematically catalog essential genes (EGs) indispensable for hPSC fitness.
- To map essential genetic determinants of hPSC fitness through genome-scale loss-of-function screens.
Main Methods:
- Performed genome-scale CRISPR loss-of-function screens.
- Utilized an inducible Cas9 H1 hPSC line.
- Cultured hPSCs on feeder cells and laminin.
Main Results:
- Identified FOXH1, VENTX, and the uncharacterized gene C22orf43/DRICH1 as essential for hPSC fitness.
- Demonstrated that hPSC EGs differ significantly from other human cell lines.
- Showed that hPSC EGs are highly context-dependent on growth substrates.
Conclusions:
- Established parameters for genome-wide screens in hPSCs.
- Facilitated the characterization of previously unappreciated genetic regulators of hPSC biology.
- Provided insights into the genetic basis of hPSC fitness and pluripotency.
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