Differentiation and CRISPR-Cas9-mediated genetic engineering of human intestinal organoids
Publication date
2022-09-16
Authors
Martinez-Silgado, Adriana
Yousef Yengej, F. A.
Puschhof, Jens
Geurts, Veerle
Boot, Charelle
Geurts, Maarten H.
Rookmaaker, Maarten B
Verhaar, Marianne C.
Beumer, Joep
Clevers, Hans
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Supervisors
Document Type
Article
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Abstract
Intestinal organoids are three-dimensional cultures that resemble key aspects of the epithelium of origin. Here, we describe how to differentiate human small intestinal organoids by combining growth media variations and genetic engineering. We detail the differentiation of human intestinal organoids in the presence and absence of BMP agonists to recapitulate a broader scope of functional cell states found in vivo. Using transient overexpression of the transcription factor Neurogenin-3, we describe the enhancement of differentiation toward rare enteroendocrine cells. For complete details on the use and execution of this protocol, please refer to Beumer et al. (2022).
Keywords
Cell differentiation, CRISPR, Organoids, CRISPR-Cas Systems/genetics, Intestines, Genetic Engineering, Humans, Cell Differentiation/genetics, General Biochemistry,Genetics and Molecular Biology, General Immunology and Microbiology, General Neuroscience, Research Support, Non-U.S. Gov't, Journal Article
Citation
Martinez-Silgado, A, Yousef Yengej, F A, Puschhof, J, Geurts, V, Boot, C, Geurts, M H, Rookmaaker, M B, Verhaar, M C, Beumer, J & Clevers, H 2022, 'Differentiation and CRISPR-Cas9-mediated genetic engineering of human intestinal organoids', STAR protocols, vol. 3, no. 3, 101639. https://doi.org/10.1016/j.xpro.2022.101639