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

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

DOI:
10.3791/64849-v

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09:52 min

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January 06, 2023

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, ,
1Obesity and Comorbidities Research Center,State University of Campinas, 2Department of Structural and Functional Biology, Institute of Biology,State University of Campinas

Chapters

  • 00:05Introduction
  • 00:53Molecular Cloning
  • 04:34In Vivo Injection of Adeno-Associated Virus (AAV) into the Inguinal White Adipose Tissue (iWAT)
  • 06:14In Vitro Differentiation of Stromal Vascular Cells (SVFs) into Beige Adipocytes
  • 07:23In Vitro AAV Infection of SVFs
  • 08:11Results: SPH-Induced Expression of Endogenous Prdm16 for Enhanced Oxygen Consumption
  • 09:23Conclusion

Summary

Automatic Translation

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

Keywords: CRISPRSunTagP65-HSF1Activation SystemAdipocytesGain-of-functionAdeno-associated VirusPRDM16SgRNACloningVirus Production

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