Generation of epitope tag knock-in mice with CRISPR-Cas9 to study the function of endogenous proteins

Zhao Zhang

doi:10.1016/j.xpro.2023.102518

Generation of epitope tag knock-in mice with CRISPR-Cas9 to study the function of endogenous proteins

Zhao Zhang

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Studying endogenous proteins in mice has provided numerous insights into the physiological and pathological roles of these proteins. However, the availability and specificity of protein-specific antibodies often limit such studies. Here we present a protocol for generating epitope tag knock-in mice with CRISPR-Cas9-mediated gene editing. We discuss key considerations for tag selection and knock-in location and provide insights into CRISPR design. Subsequently, we outline the sequential steps involved in knock-in mouse generation, genotyping, and validation and explore potential applications. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).¹

Original language	English (US)
Article number	102518
Journal	STAR Protocols
Volume	4
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2023.102518
State	Published - Sep 15 2023

Keywords

CRISPR
Genetics

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2023.102518

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abstract = "Studying endogenous proteins in mice has provided numerous insights into the physiological and pathological roles of these proteins. However, the availability and specificity of protein-specific antibodies often limit such studies. Here we present a protocol for generating epitope tag knock-in mice with CRISPR-Cas9-mediated gene editing. We discuss key considerations for tag selection and knock-in location and provide insights into CRISPR design. Subsequently, we outline the sequential steps involved in knock-in mouse generation, genotyping, and validation and explore potential applications. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).1",

keywords = "CRISPR, Genetics",

author = "Zhao Zhang",

note = "Funding Information: Research reported in this publication was supported by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health under Award Number R00DK115766 and R01DK130959 . The content is solely the responsibility of the author and does not necessarily represent the official views of the National Institutes of Health. Publisher Copyright: {\textcopyright} 2023 The Author",

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N1 - Funding Information: Research reported in this publication was supported by the National Institute of Diabetes and Digestive and Kidney Diseases of the National Institutes of Health under Award Number R00DK115766 and R01DK130959 . The content is solely the responsibility of the author and does not necessarily represent the official views of the National Institutes of Health. Publisher Copyright: © 2023 The Author

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Y1 - 2023/9/15

N2 - Studying endogenous proteins in mice has provided numerous insights into the physiological and pathological roles of these proteins. However, the availability and specificity of protein-specific antibodies often limit such studies. Here we present a protocol for generating epitope tag knock-in mice with CRISPR-Cas9-mediated gene editing. We discuss key considerations for tag selection and knock-in location and provide insights into CRISPR design. Subsequently, we outline the sequential steps involved in knock-in mouse generation, genotyping, and validation and explore potential applications. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).1

AB - Studying endogenous proteins in mice has provided numerous insights into the physiological and pathological roles of these proteins. However, the availability and specificity of protein-specific antibodies often limit such studies. Here we present a protocol for generating epitope tag knock-in mice with CRISPR-Cas9-mediated gene editing. We discuss key considerations for tag selection and knock-in location and provide insights into CRISPR design. Subsequently, we outline the sequential steps involved in knock-in mouse generation, genotyping, and validation and explore potential applications. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2022).1

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Generation of epitope tag knock-in mice with CRISPR-Cas9 to study the function of endogenous proteins

Abstract

Keywords

ASJC Scopus subject areas

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