Differentiation and CRISPR-Cas9-mediated genetic engineering of human intestinal organoids

Adriana Martinez-Silgado, Fjodor A. Yousef Yengej, Jens Puschhof, Veerle Geurts, Charelle Boot, Maarten H. Geurts, Maarten B. Rookmaaker, Marianne C. Verhaar, Joep Beumer, Hans Clevers

Onderzoeksoutput: Bijdrage aan tijdschriftArtikelpeer review

Samenvatting

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).

Originele taal-2Engels
Artikelnummer101639
TijdschriftSTAR Protocols
Volume3
Nummer van het tijdschrift3
DOI's
StatusGepubliceerd - 16 sep. 2022
Extern gepubliceerdJa

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