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Publication : Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in?vivo.

First Author  Xue K Year  2022
Journal  STAR Protoc Volume  3
Issue  4 Pages  101895
PubMed ID  36595932 Mgi Jnum  J:374089
Mgi Id  MGI:8257475 Doi  10.1016/j.xpro.2022.101895
Citation  Xue K, et al. (2022) Specific and efficient gene knockout and overexpression in mouse interscapular brown adipocytes in vivo. STAR Protoc 3(4):101895
abstractText  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).
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