Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools
Publication date
2024-09-20
Authors
Celotti, Martina
Derks, Lucca L.M.
van Es, Johan
Van Boxtel, Ruben
Clevers, Hans
Geurts, Maarten H.
Editors
Advisors
Supervisors
Document Type
Article
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License
cc_by
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
Keywords
Cancer, CRISPR, Genetics, Stem Cells, General Neuroscience, General Biochemistry,Genetics and Molecular Biology, General Immunology and Microbiology
Citation
Celotti, M, Derks, L L M, van Es, J, van Boxtel, R, Clevers, H & Geurts, M H 2024, 'Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools', STAR protocols, vol. 5, no. 3, 103189. https://doi.org/10.1016/j.xpro.2024.103189