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Cited by 55Digestive system and related healthDevelopmental Biology and Gene RegulationCRISPR and Genetic Engineering

An organoid-based CRISPR-Cas9 screen for regulators of intestinal epithelial maturation and cell fate

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Hansen, Stine L., Larsen, Hjalte L., Pikkupeura, Laura M., Maciag, Grzegorz, Guiu, Jordi, Müller, Iris, Clement, Ditte L., Mueller, Christina +4 more·2023·Science Advances
DOI 10.1126/sciadv.adg4055PMC PMC10337909
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Abstract

Generation of functionally mature organs requires exquisite control of transcriptional programs governing cell state transitions during development. Despite advances in understanding the behavior of adult intestinal stem cells and their progeny, the transcriptional regulators that control the emergence of the mature intestinal phenotype remain largely unknown. Using mouse fetal and adult small intestinal organoids, we uncover transcriptional differences between the fetal and adult state and identify rare adult-like cells present in fetal organoids. This suggests that fetal organoids have an inherent potential to mature, which is locked by a regulatory program. By implementing a CRISPR-Cas9 screen targeting transcriptional regulators expressed in fetal organoids, we establish Smarca4 and Smarcc1 as important factors safeguarding the immature progenitor state. Our approach demonstrates the utility of organoid models in the identification of factors regulating cell fate and state transitions during tissue maturation and reveals that SMARCA4 and SMARCC1 prevent precocious differentiation during intestinal development.

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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 [20]

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References 57 · Cited by 55

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Authors
Hansen, S. L., Larsen, H. L., Pikkupeura, L. M., Maciag, G., Guiu, J., Müller, I., Clement, D. L., Mueller, C., Johansen, J. V., Helin, K., Lerdrup, M., & Jensen, K. B.
Affiliations
University of Copenhagen (DK)
Corresponding author
  • Hansen, Stine L.
  • Larsen, Hjalte L.
  • Jensen, Kim B.
ORCID
Venue
Science Advances
Publisher
American Association for the Advancement of Science (AAAS)
Published
2023-07-14
Volume
9
Issue
28
Pages
eadg4055-eadg4055
ISSN
2375-2548 (ISSN-L)
DOI
10.1126/sciadv.adg4055
PMC
PMC10337909
OpenAlex
W4383988552
Cited by
55
References
57
Citations by year
2026: 15 · 2025: 21 · 2024: 16 · 2023: 3
FWCI
9.13
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Science Advances (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by)
  • Dipòsit Digital de la Universitat de Barcelona (Universitat de Barcelona) (submittedVersion, cc-by)
Funders
  • Lundbeckfonden (R303-2018-3391)
  • Novo Nordisk Fonden (NNF20OC0064376)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Digestive system and related health
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
Keywords
intestinal organoids, small intestine development, cell fate transitions, transcription factors, SMARCA4, SMARCC1, CRISPR-Cas9 screen, intestinal stem cells, chromatin remodeling
MeSH terms
Animals, Cell Differentiation, Fetus, Organoids, Mice, Adult Stem Cells, CRISPR-Cas Systems

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