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Generering af brune fedtspecifikke knockout-mus ved hjælp af et kombineret Cre-LoxP-, CRISPR-Cas9- og adeno-associeret virus-single-guide-RNA-system
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Generering af brune fedtspecifikke knockout-mus ved hjælp af et kombineret Cre-LoxP-, CRISPR-Cas9- og adeno-associeret virus-single-guide-RNA-system

Article DOI: 10.3791/65083-v • 03:58 min • March 24th, 2023
March 24th, 2023 •
1, 1, 1
1Section on Integrative Physiology and Metabolism, Research Division, Joslin Diabetes Center, Harvard Medical School

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I denne protokol beskriver vi de tekniske procedurer til generering af brune fedtvæv (BAT)-specifikke knockout-mus, der udnytter et kombineret Cre-LoxP-, CRISPR-Cas9- og adeno-associeret virus (AAV) single-guide RNA (sgRNA) system. De beskrevne trin omfatter design af sgRNA'erne, fremstilling af AAV-sgRNA-partiklerne og mikroinjektion af AAV i BAT-loberne.

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Denne måned i JoVE udgave 193 CRISPR-Cas9 AAV-sgRNA-system mus brunt fedtvæv mikroinjektion
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