Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies
Summary: Human pluripotent stem cells (hPSCs) provide an invaluable tool for modeling diseases and hold promise for regenerative medicine. For understanding pluripotency and lineage differentiation mechanisms, a critical first step involves systematically cataloging essential genes (EGs) that are in...
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| Format: | Article |
| Language: | English |
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Elsevier
2019-04-01
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| Series: | Cell Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2211124719302128 |
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doaj-b48e7be0191349729bde89c8056e0021 |
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Article |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Barbara Mair Jelena Tomic Sanna N. Masud Peter Tonge Alexander Weiss Matej Usaj Amy Hin Yan Tong Jamie J. Kwan Kevin R. Brown Emily Titus Michael Atkins Katherine S.K. Chan Lise Munsie Andrea Habsid Hong Han Marion Kennedy Brenda Cohen Gordon Keller Jason Moffat |
| spellingShingle |
Barbara Mair Jelena Tomic Sanna N. Masud Peter Tonge Alexander Weiss Matej Usaj Amy Hin Yan Tong Jamie J. Kwan Kevin R. Brown Emily Titus Michael Atkins Katherine S.K. Chan Lise Munsie Andrea Habsid Hong Han Marion Kennedy Brenda Cohen Gordon Keller Jason Moffat Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies Cell Reports |
| author_facet |
Barbara Mair Jelena Tomic Sanna N. Masud Peter Tonge Alexander Weiss Matej Usaj Amy Hin Yan Tong Jamie J. Kwan Kevin R. Brown Emily Titus Michael Atkins Katherine S.K. Chan Lise Munsie Andrea Habsid Hong Han Marion Kennedy Brenda Cohen Gordon Keller Jason Moffat |
| author_sort |
Barbara Mair |
| title |
Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies |
| title_short |
Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies |
| title_full |
Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies |
| title_fullStr |
Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies |
| title_full_unstemmed |
Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies |
| title_sort |
essential gene profiles for human pluripotent stem cells identify uncharacterized genes and substrate dependencies |
| publisher |
Elsevier |
| series |
Cell Reports |
| issn |
2211-1247 |
| publishDate |
2019-04-01 |
| description |
Summary: Human pluripotent stem cells (hPSCs) provide an invaluable tool for modeling diseases and hold promise for regenerative medicine. For understanding pluripotency and lineage differentiation mechanisms, a critical first step involves systematically cataloging essential genes (EGs) that are indispensable for hPSC fitness, defined as cell reproduction in this study. To map essential genetic determinants of hPSC fitness, we performed genome-scale loss-of-function screens in an inducible Cas9 H1 hPSC line cultured on feeder cells and laminin to identify EGs. Among these, we found FOXH1 and VENTX, genes that encode transcription factors previously implicated in stem cell biology, as well as an uncharacterized gene, C22orf43/DRICH1. hPSC EGs are substantially different from other human model cell lines, and EGs in hPSCs are highly context dependent with respect to different growth substrates. Our CRISPR screens establish parameters for genome-wide screens in hPSCs, which will facilitate the characterization of unappreciated genetic regulators of hPSC biology. : Mair et al. establish a robust, inducible CRISPR screening platform for forward genetics in human pluripotent stem cells (hPSCs). Genome-wide proliferation screens identified core essential genes for hPSCs and revealed context-dependent genetic requirements on different substrates. This underlines hPSC plasticity and helps us to understand the genetic wiring of hPSCs. Keywords: human pluripotent stem cells, genome-wide CRISPR screen, functional genomics, essential genes, DRICH1 |
| url |
http://www.sciencedirect.com/science/article/pii/S2211124719302128 |
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doaj-b48e7be0191349729bde89c8056e00212020-11-25T02:07:09ZengElsevierCell Reports2211-12472019-04-01272599615.e12Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate DependenciesBarbara Mair0Jelena Tomic1Sanna N. Masud2Peter Tonge3Alexander Weiss4Matej Usaj5Amy Hin Yan Tong6Jamie J. Kwan7Kevin R. Brown8Emily Titus9Michael Atkins10Katherine S.K. Chan11Lise Munsie12Andrea Habsid13Hong Han14Marion Kennedy15Brenda Cohen16Gordon Keller17Jason Moffat18Donnelly Centre, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON, CanadaCentre for Commercialization of Regenerative Medicine, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaMcEwen Stem Cell Institute, University Health Network, University of Toronto, Toronto, ON, Canada; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaCentre for Commercialization of Regenerative Medicine, Toronto, ON, CanadaMcEwen Stem Cell Institute, University Health Network, University of Toronto, Toronto, ON, Canada; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaCentre for Commercialization of Regenerative Medicine, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, CanadaMcEwen Stem Cell Institute, University Health Network, University of Toronto, Toronto, ON, Canada; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, CanadaMcEwen Stem Cell Institute, University Health Network, University of Toronto, Toronto, ON, Canada; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, CanadaMcEwen Stem Cell Institute, University Health Network, University of Toronto, Toronto, ON, Canada; Princess Margaret Cancer Centre, University Health Network, University of Toronto, Toronto, ON, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON, CanadaDonnelly Centre, University of Toronto, Toronto, ON, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada; Canadian Institute for Advanced Research, Toronto, ON, Canada; Institute for Biomaterials and BioMedical Engineering, University of Toronto, ON, Canada; Corresponding authorSummary: Human pluripotent stem cells (hPSCs) provide an invaluable tool for modeling diseases and hold promise for regenerative medicine. For understanding pluripotency and lineage differentiation mechanisms, a critical first step involves systematically cataloging essential genes (EGs) that are indispensable for hPSC fitness, defined as cell reproduction in this study. To map essential genetic determinants of hPSC fitness, we performed genome-scale loss-of-function screens in an inducible Cas9 H1 hPSC line cultured on feeder cells and laminin to identify EGs. Among these, we found FOXH1 and VENTX, genes that encode transcription factors previously implicated in stem cell biology, as well as an uncharacterized gene, C22orf43/DRICH1. hPSC EGs are substantially different from other human model cell lines, and EGs in hPSCs are highly context dependent with respect to different growth substrates. Our CRISPR screens establish parameters for genome-wide screens in hPSCs, which will facilitate the characterization of unappreciated genetic regulators of hPSC biology. : Mair et al. establish a robust, inducible CRISPR screening platform for forward genetics in human pluripotent stem cells (hPSCs). Genome-wide proliferation screens identified core essential genes for hPSCs and revealed context-dependent genetic requirements on different substrates. This underlines hPSC plasticity and helps us to understand the genetic wiring of hPSCs. Keywords: human pluripotent stem cells, genome-wide CRISPR screen, functional genomics, essential genes, DRICH1http://www.sciencedirect.com/science/article/pii/S2211124719302128 |