Rapid establishment of human colonic organoid knockout lines.

Abstract

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).

• Authors: Wei Gu, Jonathan L Colarusso, Michael K Dame, Jason R Spence, Qiao Zhou
• Journal: STAR protocols (STAR Protoc) Vol. 3 Issue 2 Pg. 101308 (06 17 2022) ISSN: 2666-1667 [Electronic] United States
• PMID: 35463469 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: © 2022 The Author(s).

Topics

• Biopsy
• Colon
• Humans
• Organoids