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

doi:10.1016/j.xpro.2022.101639

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

Hubrecht Institute

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgave › Artikel › Wetenschappelijk › peer review

4 Citaten (Scopus)

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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-2	Engels
Pagina's (van-tot)	101639
Tijdschrift	STAR protocols
Volume	3
Nummer van het tijdschrift	3
DOI's	https://doi.org/10.1016/j.xpro.2022.101639
Status	Gepubliceerd - 16 sep. 2022

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10.1016/j.xpro.2022.101639

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title = "Differentiation and CRISPR-Cas9-mediated genetic engineering of human intestinal organoids",

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 = "CRISPR-Cas Systems/genetics, Cell Differentiation/genetics, Genetic Engineering, Humans, Intestines, Organoids",

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year = "2022",

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doi = "10.1016/j.xpro.2022.101639",

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T1 - Differentiation and CRISPR-Cas9-mediated genetic engineering of human intestinal organoids

AU - Martinez-Silgado, Adriana

AU - Yousef Yengej, Fjodor A

AU - Puschhof, Jens

AU - Geurts, Veerle

AU - Boot, Charelle

AU - Geurts, Maarten H

AU - Rookmaaker, Maarten B

AU - Verhaar, Marianne C

AU - Beumer, Joep

AU - Clevers, Hans

PY - 2022/9/16

Y1 - 2022/9/16

N2 - 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).

AB - 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).

KW - CRISPR-Cas Systems/genetics

KW - Cell Differentiation/genetics

KW - Genetic Engineering

KW - Humans

KW - Intestines

KW - Organoids

U2 - 10.1016/j.xpro.2022.101639

DO - 10.1016/j.xpro.2022.101639

M3 - Article

C2 - 36042877

SN - 2666-1667

VL - 3

SP - 101639

JO - STAR protocols

JF - STAR protocols

IS - 3

ER -

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

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