State University of Campinas 1 article published in JoVE Bioengineering Investigation of Beige Fat Biology and Metabolism Using the CRISPR SunTag-p65-HSF1 Activation System Fernando Bladimir Valdivieso-Rivera*1, Vanessa de Oliveira Furino*1, Carlos Henrique Sponton1,2 1Obesity and Comorbidities Research Center, State University of Campinas, 2Department of Structural and Functional Biology, Institute of Biology, State University of Campinas 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.