Protocol for targeted gene manipulation and thermogenic evaluation in mouse brown adipocytes.

Adeno-associated viruses (AAVs) are versatile, non-integrating vectors for in vivo gene delivery. We present a reproducible workflow for generating Cre-dependent FLEX-AAVs, quantifying viral titer, and performing localized injections for cell-type-specific transgene expression in mice. The protocol also details the assessment of thermogenic capacity in genetically modified brown adipocytes using Clark-type electrode respirometry. For complete details on the use and execution of this protocol, please refer to Bunk et al.1.