Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools

Martina Celotti; Lucca L M Derks; Johan van Es; Ruben van Boxtel; Hans Clevers; Maarten H Geurts

doi:10.1016/j.xpro.2024.103189

Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools

Martina Celotti, Lucca L M Derks, Johan van Es, Ruben van Boxtel, Hans Clevers, Maarten H Geurts

Hubrecht Institute

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

4 Citaten (Scopus)

Samenvatting

Isogenic disease models, such as genetically engineered organoids, provide insight into the impact of genetic variants on organ function. Here, we present a protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools. We describe steps for single guide RNA (sgRNA) design and cloning, electroporation, and selecting electroporated cells. We then detail procedures for clonal line generation. Next-generation CRISPR tools do not require double-stranded break (DSB) induction for their function, thus simplifying in vitro disease model generation. For complete details on the use and execution of this protocol, please refer to Geurts et al.1,2.

Originele taal-2	Engels
Pagina's (van-tot)	103189
Tijdschrift	STAR protocols
Volume	5
Nummer van het tijdschrift	3
DOI's	https://doi.org/10.1016/j.xpro.2024.103189
Status	Gepubliceerd - 20 sep. 2024

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10.1016/j.xpro.2024.103189

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title = "Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools",

abstract = "Isogenic disease models, such as genetically engineered organoids, provide insight into the impact of genetic variants on organ function. Here, we present a protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools. We describe steps for single guide RNA (sgRNA) design and cloning, electroporation, and selecting electroporated cells. We then detail procedures for clonal line generation. Next-generation CRISPR tools do not require double-stranded break (DSB) induction for their function, thus simplifying in vitro disease model generation. For complete details on the use and execution of this protocol, please refer to Geurts et al.1,2.",

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AU - Clevers, Hans

AU - Geurts, Maarten H

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KW - Gene Editing/methods

KW - Clustered Regularly Interspaced Short Palindromic Repeats/genetics

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