Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology

We provide a protocol for gene editing in mouse hepatocytes in vivo using the CRISPR-Cas9 technology via AAV delivery. This protocol describes the construction of AAV plasmids, AAV packaging, injection, and the detection of in vivo knockout efficiency. Using this protocol, we can get up to 10(14) AA...

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Detalles Bibliográficos
Autores principales: Chen, Yanhao, Ding, Qiurong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715323/
https://www.ncbi.nlm.nih.gov/pubmed/35005644
http://dx.doi.org/10.1016/j.xpro.2021.101062
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author Chen, Yanhao
Ding, Qiurong
author_facet Chen, Yanhao
Ding, Qiurong
author_sort Chen, Yanhao
collection PubMed
description We provide a protocol for gene editing in mouse hepatocytes in vivo using the CRISPR-Cas9 technology via AAV delivery. This protocol describes the construction of AAV plasmids, AAV packaging, injection, and the detection of in vivo knockout efficiency. Using this protocol, we can get up to 10(14) AAV and knock out genes in hepatocytes efficiently within 15 days. Moreover, we describe an optimized protocol to simultaneously target two genes via AAV delivery of CRISPR-Cas9 materials in the liver. For complete details on the use and execution of this profile, please refer to Wei et al. (2020).
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spelling pubmed-87153232022-01-06 Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology Chen, Yanhao Ding, Qiurong STAR Protoc Protocol We provide a protocol for gene editing in mouse hepatocytes in vivo using the CRISPR-Cas9 technology via AAV delivery. This protocol describes the construction of AAV plasmids, AAV packaging, injection, and the detection of in vivo knockout efficiency. Using this protocol, we can get up to 10(14) AAV and knock out genes in hepatocytes efficiently within 15 days. Moreover, we describe an optimized protocol to simultaneously target two genes via AAV delivery of CRISPR-Cas9 materials in the liver. For complete details on the use and execution of this profile, please refer to Wei et al. (2020). Elsevier 2021-12-23 /pmc/articles/PMC8715323/ /pubmed/35005644 http://dx.doi.org/10.1016/j.xpro.2021.101062 Text en © 2021 The Authors 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
Chen, Yanhao
Ding, Qiurong
Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology
title Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology
title_full Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology
title_fullStr Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology
title_full_unstemmed Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology
title_short Optimized protocols for efficient gene editing in mouse hepatocytes in vivo using CRISPR-Cas9 technology
title_sort optimized protocols for efficient gene editing in mouse hepatocytes in vivo using crispr-cas9 technology
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8715323/
https://www.ncbi.nlm.nih.gov/pubmed/35005644
http://dx.doi.org/10.1016/j.xpro.2021.101062
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AT dingqiurong optimizedprotocolsforefficientgeneeditinginmousehepatocytesinvivousingcrisprcas9technology

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