Rapid establishment of human colonic organoid knockout lines

Human colonic organoids derived from biopsy or autopsy tissues are a vital tool to study mucosal homeostasis, model colonic diseases, and develop therapeutics. Rapid and reliable generation of knockout organoid lines from multiple donors enables analysis of specific gene functions. Here, we report p...

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Detalles Bibliográficos
Autores principales: Gu, Wei, Colarusso, Jonathan L., Dame, Michael K., Spence, Jason R., Zhou, Qiao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9026586/
https://www.ncbi.nlm.nih.gov/pubmed/35463469
http://dx.doi.org/10.1016/j.xpro.2022.101308
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author Gu, Wei
Colarusso, Jonathan L.
Dame, Michael K.
Spence, Jason R.
Zhou, Qiao
author_facet Gu, Wei
Colarusso, Jonathan L.
Dame, Michael K.
Spence, Jason R.
Zhou, Qiao
author_sort Gu, Wei
collection PubMed
description Human colonic organoids derived from biopsy or autopsy tissues are a vital tool to study mucosal homeostasis, model colonic diseases, and develop therapeutics. Rapid and reliable generation of knockout organoid lines from multiple donors enables analysis of specific gene functions. Here, we report protocols to produce colonic organoid knockout lines within 1 to 2 weeks using lentiviral delivery of CRISPR-Cas9, achieving knockout efficiency of 90% or greater. These lines are suitable for multi-lineage differentiation and downstream analysis. For complete details on the use and execution of this protocol, please refer to Gu et al. (2022).
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spelling pubmed-90265862022-04-23 Rapid establishment of human colonic organoid knockout lines Gu, Wei Colarusso, Jonathan L. Dame, Michael K. Spence, Jason R. Zhou, Qiao STAR Protoc Protocol Human colonic organoids derived from biopsy or autopsy tissues are a vital tool to study mucosal homeostasis, model colonic diseases, and develop therapeutics. Rapid and reliable generation of knockout organoid lines from multiple donors enables analysis of specific gene functions. Here, we report protocols to produce colonic organoid knockout lines within 1 to 2 weeks using lentiviral delivery of CRISPR-Cas9, achieving knockout efficiency of 90% or greater. These lines are suitable for multi-lineage differentiation and downstream analysis. For complete details on the use and execution of this protocol, please refer to Gu et al. (2022). Elsevier 2022-04-12 /pmc/articles/PMC9026586/ /pubmed/35463469 http://dx.doi.org/10.1016/j.xpro.2022.101308 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Gu, Wei
Colarusso, Jonathan L.
Dame, Michael K.
Spence, Jason R.
Zhou, Qiao
Rapid establishment of human colonic organoid knockout lines
title Rapid establishment of human colonic organoid knockout lines
title_full Rapid establishment of human colonic organoid knockout lines
title_fullStr Rapid establishment of human colonic organoid knockout lines
title_full_unstemmed Rapid establishment of human colonic organoid knockout lines
title_short Rapid establishment of human colonic organoid knockout lines
title_sort rapid establishment of human colonic organoid knockout lines
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9026586/
https://www.ncbi.nlm.nih.gov/pubmed/35463469
http://dx.doi.org/10.1016/j.xpro.2022.101308
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