Targeted ablation of Lgr5-expressing intestinal stem cells in diphtheria toxin receptor-based mouse and organoid models

Hui Yi Grace Lim; Swathi Yada; Nick Barker

doi:10.1016/j.xpro.2022.101411

Targeted ablation of Lgr5-expressing intestinal stem cells in diphtheria toxin receptor-based mouse and organoid models

STAR Protoc. 2022 May 18;3(2):101411. doi: 10.1016/j.xpro.2022.101411. eCollection 2022 Jun 17.

Authors

Hui Yi Grace Lim¹, Swathi Yada¹, Nick Barker^{1

2}

Affiliations

¹ A∗STAR Institute of Molecular and Cell Biology, Singapore 138648, Singapore.
² Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117593, Singapore.

Abstract

Intestinal cells marked by Lgr5 function as tissue-resident stem cells that sustain the homeostatic replenishment of the epithelium. By incorporating a diphtheria toxin receptor (DTR) cassette linked to the Lgr5 coding region, native Lgr5-expressing cells are susceptible to ablation upon DT administration in vivo. A similar strategy can be used for Lgr5-expressing cells within organoids established from DTR models. Together, these in vivo and in vitro approaches will facilitate dissection of the roles of Lgr5-expressing cells residing in different tissue compartments. For complete details on the use and execution of this protocol, please refer to Tan et al. (2021).

Keywords: Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Heparin-binding EGF-like Growth Factor / genetics
Intestines
Mice
Organoids*
Receptors, G-Protein-Coupled* / genetics
Stem Cells

Substances

Heparin-binding EGF-like Growth Factor
Lgr5 protein, mouse
Receptors, G-Protein-Coupled