Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system.

TitleGeneration of knock-in degron tags for endogenous proteins in mice using the dTAG system.
Publication TypeJournal Article
Year of Publication2022
AuthorsAbuhashem A, Hadjantonakis A-K
JournalSTAR Protoc
Volume3
Issue3
Pagination101660
Date Published2022 Sep 16
ISSN2666-1667
KeywordsAnimals, Chimera, Mice, Proteins, Research
Abstract

Controlling the abundance of a protein of interest in vivo is crucial to study its function. Here, we provide a step-by-step protocol for generating genetically engineered mouse (GEM) models harboring a degradation tag (dTAG) fused to endogenous proteins to enable their degradation. We discuss considerations for the overall design and details for vectors generation. Then, we include steps for generation and validations of edited mouse embryonic stem cells followed by mouse colony establishment via chimeric mouse generation. For complete details on the use and execution of this protocol, please refer to Abuhashem et al. (2022c).

DOI10.1016/j.xpro.2022.101660
Alternate JournalSTAR Protoc
PubMed ID36097386
PubMed Central IDPMC9467879
Grant ListF30 HD103398 / HD / NICHD NIH HHS / United States

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