Submitted by bel2021 on September 23, 2022 - 3:37pm
Title | Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system. |
Publication Type | Journal Article |
Year of Publication | 2022 |
Authors | Abuhashem A, Hadjantonakis A-K |
Journal | STAR Protoc |
Volume | 3 |
Issue | 3 |
Pagination | 101660 |
Date Published | 2022 Sep 16 |
ISSN | 2666-1667 |
Keywords | Animals, 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). |
DOI | 10.1016/j.xpro.2022.101660 |
Alternate Journal | STAR Protoc |
PubMed ID | 36097386 |
PubMed Central ID | PMC9467879 |
Grant List | F30 HD103398 / HD / NICHD NIH HHS / United States |