气候变化领域集成服务门户(CCPortal): A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells

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DOI	10.1073/pnas.2016806118
	A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells
	Murakami K.; Terakado Y.; Saito K.; Jomen Y.; Takeda H.; Oshima M.; Barker N.
发表日期	2021
ISSN	00278424
卷号	118 期号:4
英文摘要	An ability to safely harness the powerful regenerative potential of adult stem cells for clinical applications is critically dependent on a comprehensive understanding of the underlying mechanisms regulating their activity. Epithelial organoid cultures accurately recapitulate many features of in vivo stem cell-driven epithelial renewal, providing an excellent ex vivo platform for interrogation of key regulatory mechanisms. Here, we employed a genome-scale clustered, regularly interspaced, short palindromic repeats (CRISPR) knockout (KO) screening assay using mouse gastric epithelial organoids to identify modulators of Wnt-driven stem cell-dependent epithelial renewal in the gastric mucosa. In addition to known Wnt pathway regulators, such as Apc, we found that KO of Alk, Bclaf3, or Prkra supports the Wnt independent self-renewal of gastric epithelial cells ex vivo. In adult mice, expression of these factors is predominantly restricted to non-Lgr5–expressing stem cell zones above the gland base, implicating a critical role for these factors in suppressing self-renewal or promoting differentiation of gastric epithelia. Notably, we found that Alk inhibits Wnt signaling by phosphorylating the tyrosine of Gsk3β, while Bclaf3 and Prkra suppress regenerating islet-derived (Reg) genes by regulating the expression of epithelial interleukins. Therefore, Alk, Bclaf3, and Prkra may suppress stemness/proliferation and function as novel regulators of gastric epithelial differentiation. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	CRISPR-Cas9; Genome-wide screening; Lgr5; Tissue stem cells; Wnt
语种	英语
scopus关键词	APC protein; glycogen synthase kinase 3beta; Wnt protein; adult; Alk gene; animal cell; animal experiment; animal genetics; animal tissue; Apc gene; Article; Bclaf3 gene; cell differentiation; cell function; cell proliferation; cell renewal; clustered regularly interspaced short palindromic repeat; colonoid; controlled study; ex vivo study; gene cluster; gene control; gene expression; gene repression; genetic screening; genome-wide association study; in vivo study; Lgr5 gene; mouse; nonhuman; priority journal; Prkra gene; protein phosphorylation; Reg gene; stem cell; stomach epithelium; stomach mucosa; Wnt signaling
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180902
作者单位	Division of Epithelial Stem Cell Biology, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa, 924-1192, Japan; Laboratory of Molecular Genetics, National Cancer Center Research Institute, Chuo-ku, Tokyo 104-0045, Japan; Division of Genetics, Cancer Research Institute, Nano-Life Science Institute, Kanazawa University, Kakuma-machi, Kanazawa, 920-1192, Japan; Epithelial Stem Cell Group, Agency for Science, Technology and Research Institute of Molecular and Cell Biology, Singapore, 138673, Singapore; Agency for Science,, Technology and Research Institute of Medical Biology, Singapore, 276543, Singapore
推荐引用方式 GB/T 7714	Murakami K.,Terakado Y.,Saito K.,et al. A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells[J],2021,118(4).
APA	Murakami K..,Terakado Y..,Saito K..,Jomen Y..,Takeda H..,...&Barker N..(2021).A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells.Proceedings of the National Academy of Sciences of the United States of America,118(4).
MLA	Murakami K.,et al."A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells".Proceedings of the National Academy of Sciences of the United States of America 118.4(2021).