Department of Internal Medicine, University of Michigan, Ann Arbor, United States
Jine Wang
Department of Internal Medicine, University of Michigan, Ann Arbor, United States; Chinese academy, College of Medical Science, China Three Gorges University, Yichang, China
Department of Internal Medicine, University of Michigan, Ann Arbor, United States
Alison H Affinati
Department of Internal Medicine, University of Michigan, Ann Arbor, United States
Jonathan N Flak
Indiana Biosciences Research Institute, Indianapolis, United States
Chien Li
Novo Nordisk Research Center, Seattle, United States
David P Olson
Department of Pediatrics, University of Michigan, Ann Arbor, United States; Department of Molecular and Integrative Physiology, Ann Arbor, United States
Department of Internal Medicine, University of Michigan, Ann Arbor, United States; Department of Molecular and Integrative Physiology, Ann Arbor, United States
While Cre-dependent viral systems permit the manipulation of many neuron types, some cell populations cannot be targeted by a single DNA recombinase. Although the combined use of Flp and Cre recombinases can overcome this limitation, insufficient recombinase activity can reduce the efficacy of existing Cre+Flp-dependent viral systems. We developed a sensitive dual recombinase-activated viral approach: tTA-driven Recombinase-Guided Intersectional Targeting (tTARGIT) adeno-associated viruses (AAVs). tTARGIT AAVs utilize a Flp-dependent tetracycline transactivator (tTA) ‘Driver’ AAV and a tetracycline response element-driven, Cre-dependent ‘Payload’ AAV to express the transgene of interest. We employed this system in Slc17a6FlpO;LeprCre mice to manipulate LepRb neurons of the ventromedial hypothalamus (VMH; LepRbVMH neurons) while omitting neighboring LepRb populations. We defined the circuitry of LepRbVMH neurons and roles for these cells in the control of food intake and energy expenditure. Thus, the tTARGIT system mediates robust recombinase-sensitive transgene expression, permitting the precise manipulation of previously intractable neural populations.
