A genome-scale CRISPR screen reveals factors regulating Wnt-dependent renewal of mouse gastric epithelial cells
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Significance: Wnt activation is critically important during both early development and throughout adulthood for the maintenance and regeneration of tissue-resident stem cells. However, the underlying mechanism regulating Wnt-dependent epithelial renewal and differentiation in the stomach remains poorly understood. Here we apply genome-scale CRISPR-Cas9 knockout screening to three-dimensional ex vivo stomach organoids to reveal important mechanistic insight into Wnt-dependent epithelial renewal in mouse gastric epithelia. Our organoid-based screening strategy can be readily applied to a wide variety of organoids derived from different sources, potentially providing important insights on multiple biological phenomena.
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CRISPR cutting screens in 3D organoids have been implemented to explore factors such as growth factor dependency for cell growth, identify potential cancer-driving forces, and uncover key genetic events in cell differentiation.
“Even more exciting is the implementation of CRISPR cutting screens in 3D organoids to explore factors such as growth factor dependency for cell growth14–16, identifying potential cancer-driving forces17–19, and uncovering key genetic events in cell differentiation20–22.”
From Large-scale CRISPR screening in primary human 3D gastric organoids enables comprehensive dissection of gene-drug interactions (2025) · cites this paper as [14]
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Gastric cancer organoids and their convergence in engineering approaches
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Random clonal expansion as a limiting factor in transplantable in vivo CRISPR/Cas9 screensPreprint
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Organoid Perturbations as Tools to Explore Cellular Function and Plasticity
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- Authors
- Murakami, K., Terakado, Y., Saito, K., Jomen, Y., Takeda, H., Oshima, M., & Barker, N.
- Affiliations
- Kanazawa University (JP)
- National Cancer Center Research Institute (JP)
- Corresponding author
- Murakami, Kazuhiro
- Barker, Nick
- ORCID
- Venue
- Proceedings of the National Academy of Sciences
- Publisher
- National Academy of Sciences
- Published
- 2021-01-21
- Volume
- 118
- Issue
- 4
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- PMC
- PMC7848749
- OpenAlex
- W3124533528
- Cited by
- 62
- References
- 42
- Citations by year
- 2026: 3 · 2025: 18 · 2024: 9 · 2023: 12 · 2022: 13 · 2021: 7
- FWCI
- 3.22
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Proceedings of the National Academy of Sciences (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- MEXT | Japan Society for the Promotion of Science (17K07161, 17H06710, 17H03586, 17H01399)
- National Research Foundation Singapore (NRF-NRF12017-03)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Health Sciences › Medicine › Oncology › Cancer Cells and Metastasis
- Keywords
- Wnt signaling, gastric epithelial cells, epithelial regeneration, gastric stem cells, CRISPR-Cas9 knockout screen, tissue regeneration
- MeSH terms
- Glycogen Synthase Kinase 3 beta, Gene Editing, Anaplastic Lymphoma Kinase, Animals, Epithelial Cells, Gastric Mucosa, Gene Expression Regulation, Humans, Interleukins, Organoids, Stomach, Gene Library, RNA-Binding Proteins, Adenomatous Polyposis Coli Protein, Receptors, G-Protein-Coupled, Cell Proliferation, Mice, Adult Stem Cells, HEK293 Cells, Wnt Signaling Pathway, CRISPR-Cas Systems
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