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JoVE Journal Bioengineering
Investigation of Beige Fat Biology and Metabolism Using the CRISPR SunTag-p65-HSF1 Activation System
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Investigation of Beige Fat Biology and Metabolism Using the CRISPR SunTag-p65-HSF1 Activation System

Article DOI: 10.3791/64849-v • 09:52 min • January 6th, 2023
January 6th, 2023 •
*1, *1, 1,2
1Obesity and Comorbidities Research Center, State University of Campinas, 2Department of Structural and Functional Biology, Institute of Biology, State University of Campinas
* These authors contributed equally

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Summary
Automatic Translation

This protocol presents the use of CRISPR SunTag-p65-HSF1 (SPH) in adipocytes (AdipoSPH) as an alternative strategy to adeno-associated virus (AAV) for investigating beige fat biology. In vivo injection of AAV-carrying sgRNA targeting the endogenous Prdm16 gene is sufficient to induce beige fat development and enhance the thermogenic gene program.

Tags

Bioengineering CRISPR SunTag-p65-HSF1 Activation System Gain Function Assays Viral Vectors Adipocytes Adipose Tissue Single Guide RNA (sgRNA) Adeno-associated Virus (AAV) Cloning Injection PRDM16 Gene CRISPR/Cas9 Annealing Oligonucleotides Sac1 Restriction Site Vector Backbone PAAVU6GRNACBHM Cherry
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