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

Research output: Contribution to journal/periodical › Article › Scientific › peer-review

4 Citations (Scopus)

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.

Original language	English
Pages (from-to)	103189
Journal	STAR protocols
Volume	5
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2024.103189
Publication status	Published - 20 Sept 2024

Keywords

Organoids/cytology
Adult Stem Cells/cytology
Humans
CRISPR-Cas Systems/genetics
Electroporation/methods
RNA, Guide, CRISPR-Cas Systems/genetics
Gene Editing/methods
Clustered Regularly Interspaced Short Palindromic Repeats/genetics

Access to Document

10.1016/j.xpro.2024.103189

Cite this

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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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year = "2024",

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doi = "10.1016/j.xpro.2024.103189",

language = "English",

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T1 - Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools

AU - Celotti, Martina

AU - Derks, Lucca L M

AU - van Es, Johan

AU - van Boxtel, Ruben

AU - Clevers, Hans

AU - Geurts, Maarten H

PY - 2024/9/20

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KW - Organoids/cytology

KW - Adult Stem Cells/cytology

KW - Humans

KW - CRISPR-Cas Systems/genetics

KW - Electroporation/methods

KW - RNA, Guide, CRISPR-Cas Systems/genetics

KW - Gene Editing/methods

KW - Clustered Regularly Interspaced Short Palindromic Repeats/genetics

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DO - 10.1016/j.xpro.2024.103189

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