Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies
| Autor: | Michael Atkins, Marion Kennedy, Kevin R. Brown, Sanna Masud, Katherine S. K. Chan, Lise Munsie, Matej Usaj, Gordon Keller, Brenda Cohen, Amy Hin Yan Tong, Barbara Mair, Alexander Weiss, Emily Titus, Andrea Habsid, Jamie J. Kwan, Jason Moffat, Hong Han, Peter Tonge, Jelena Tomic |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Pluripotent Stem Cells Genes Essential Cas9 Context (language use) Cell Differentiation Computational biology Biology Regenerative medicine General Biochemistry Genetics and Molecular Biology 3. Good health 03 medical and health sciences 030104 developmental biology 0302 clinical medicine lcsh:Biology (General) Gene Expression Regulation Essential gene CRISPR Humans Clustered Regularly Interspaced Short Palindromic Repeats Stem cell Induced pluripotent stem cell lcsh:QH301-705.5 Functional genomics 030217 neurology & neurosurgery |
| Zdroj: | Cell Reports, Vol 27, Iss 2, Pp 599-615.e12 (2019) |
| ISSN: | 2211-1247 |
| Popis: | 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 |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|