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Title: Generation of gene-of-interest knockouts in murine organoids using CRISPR-Cas9.
Austin Authors: Huber, Anne;Dijkstra, Christine ;Ernst, Matthias ;Eissmann, Moritz F 
Affiliation: Olivia Newton-John Cancer Research Institute
La Trobe School of Cancer Medicine, La Trobe University, Melbourne, VIC 3084, Australia.
Issue Date: 30-Jan-2023 2023
Publication information: STAR Protocols 2023; 4(1)
Abstract: Gene-of-interest knockout organoids present a powerful and versatile research tool to study a gene's effects on many biological and pathological processes. Here, we present a straightforward and broadly applicable protocol to generate gene knockouts in mouse organoids using CRISPR-Cas9 technology. We describe the processes of transient transfecting organoids with pre-assembled CRISPR-Cas9 ribonucleoprotein complexes, organoid cell sorting, and establishing clonal organoid culture pairs. We then detail how to confirm the knockout via Western blot analysis.
DOI: 10.1016/j.xpro.2023.102076
Journal: STAR Protocols
Start page: 102076
PubMed URL: 36853714
ISSN: 2666-1667
Type: Journal Article
Subjects: CRISPR
Appears in Collections:Journal articles

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