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. 2022 Oct 27;12:18051. doi: 10.1038/s41598-022-21706-2

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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CMT organoids can be genetically modified with the CRISPR/Cas9 system. (a) GFP was introduced into ORG by lentiviral transduction at the indicated multiplicities of infection (MOI). GFP expression was analyzed by flow cytometry 3 days after transduction. For MOI 1, Mean ± SD = 12.6 ± 4.2%, for MOI 4, Mean ± SD = 35.3 ± 6.6%. Two independent experiments that were conducted with three different organoid lines are presented, demonstrating robust transduction efficiency. (b) Frequency of frameshift indels in organoids ORG-25-C1 modified by CRISPR/Cas9 using a gRNA targeting vimentin (gVIM-1 and gVIM-2), compared to the non-targeting (NT) gRNA. TIDE analysis two passages after transduction. (c) Representative images of H&E stainings and immunohistochemistry of VIM in organoids ORG-25-C1 modified by CRISPR/Cas9 using a gRNA targeting vimentin (gVIM-1 and gVIM-2), compared to the non-targeting (NT) gRNA. Embedding four passages after transduction. Scale bar, 50 μm.