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Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo
The classical Cre-LoxP system is time consuming. Here we detail a protocol that leverages Rosa26-LSL-Cas9;Adiponectin-Cre mice to restrict Cas9 expression in adipocytes. This enables specific deletion of target genes in brown adipocytes within 6 weeks by local injection of AAV-sgRNA into interscapul...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9722717/ https://www.ncbi.nlm.nih.gov/pubmed/36595932 http://dx.doi.org/10.1016/j.xpro.2022.101895 |
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author | Xue, Kaili Wu, Dongmei Qiu, Yifu |
author_facet | Xue, Kaili Wu, Dongmei Qiu, Yifu |
author_sort | Xue, Kaili |
collection | PubMed |
description | The classical Cre-LoxP system is time consuming. Here we detail a protocol that leverages Rosa26-LSL-Cas9;Adiponectin-Cre mice to restrict Cas9 expression in adipocytes. This enables specific deletion of target genes in brown adipocytes within 6 weeks by local injection of AAV-sgRNA into interscapular brown adipose tissue. We also describe an adiponectin-promoter-driven AAV vector to express sgRNA-resistant cDNA-encoded protein for subsequent rescue. This protocol thus provides an efficient means to specifically knockout and overexpress genes in brown adipocytes in vivo. For complete details on the use and execution of this protocol, please refer to Xue et al. (2022).(1) |
format | Online Article Text |
id | pubmed-9722717 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-97227172022-12-07 Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo Xue, Kaili Wu, Dongmei Qiu, Yifu STAR Protoc Protocol The classical Cre-LoxP system is time consuming. Here we detail a protocol that leverages Rosa26-LSL-Cas9;Adiponectin-Cre mice to restrict Cas9 expression in adipocytes. This enables specific deletion of target genes in brown adipocytes within 6 weeks by local injection of AAV-sgRNA into interscapular brown adipose tissue. We also describe an adiponectin-promoter-driven AAV vector to express sgRNA-resistant cDNA-encoded protein for subsequent rescue. This protocol thus provides an efficient means to specifically knockout and overexpress genes in brown adipocytes in vivo. For complete details on the use and execution of this protocol, please refer to Xue et al. (2022).(1) Elsevier 2022-12-02 /pmc/articles/PMC9722717/ /pubmed/36595932 http://dx.doi.org/10.1016/j.xpro.2022.101895 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 Xue, Kaili Wu, Dongmei Qiu, Yifu Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
title | Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
title_full | Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
title_fullStr | Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
title_full_unstemmed | Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
title_short | Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
title_sort | specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9722717/ https://www.ncbi.nlm.nih.gov/pubmed/36595932 http://dx.doi.org/10.1016/j.xpro.2022.101895 |
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