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

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 consi...

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Detalles Bibliográficos
Autores principales: Abuhashem, Abderhman, Hadjantonakis, Anna-Katerina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9467879/
https://www.ncbi.nlm.nih.gov/pubmed/36097386
http://dx.doi.org/10.1016/j.xpro.2022.101660
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author Abuhashem, Abderhman
Hadjantonakis, Anna-Katerina
author_facet Abuhashem, Abderhman
Hadjantonakis, Anna-Katerina
author_sort Abuhashem, Abderhman
collection PubMed
description 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).
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spelling pubmed-94678792022-09-14 Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system Abuhashem, Abderhman Hadjantonakis, Anna-Katerina STAR Protoc Protocol 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). Elsevier 2022-09-07 /pmc/articles/PMC9467879/ /pubmed/36097386 http://dx.doi.org/10.1016/j.xpro.2022.101660 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Abuhashem, Abderhman
Hadjantonakis, Anna-Katerina
Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system
title Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system
title_full Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system
title_fullStr Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system
title_full_unstemmed Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system
title_short Generation of knock-in degron tags for endogenous proteins in mice using the dTAG system
title_sort generation of knock-in degron tags for endogenous proteins in mice using the dtag system
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9467879/
https://www.ncbi.nlm.nih.gov/pubmed/36097386
http://dx.doi.org/10.1016/j.xpro.2022.101660
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