Authors: | Abuhashem, A.; Hadjantonakis, A. K. |
Article Title: | Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system |
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). © 2022 The Author(s) |
Keywords: | genetics; microscopy; proteins; mouse; animal; metabolism; animals; mice; protein; developmental biology; chimera; cell culture; research; stem cells; molecular biology; model organisms; crispr |
Journal Title: | STAR Protocols |
Volume: | 3 |
Issue: | 3 |
ISSN: | 2666-1667 |
Publisher: | Cell Press |
Date Published: | 2022-09-16 |
Start Page: | 101660 |
Language: | English |
DOI: | 10.1016/j.xpro.2022.101660 |
PUBMED: | 36097386 |
PROVIDER: | scopus |
PMCID: | PMC9467879 |
DOI/URL: | |
Notes: | Article -- Export Date: 3 October 2022 -- Source: Scopus |