tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice.
Paul V SabatiniJine WangAlan C RuppAlison H AffinatiJonathan N FlakChien LiDavid P OlsonMartin G MyersPublished in: eLife (2021)
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.
Keyphrases
- sars cov
- spinal cord
- genetic diversity
- induced apoptosis
- cancer therapy
- single cell
- stem cells
- high fat diet induced
- metabolic syndrome
- type diabetes
- oxidative stress
- cell cycle arrest
- drug delivery
- cell therapy
- circulating tumor
- blood brain barrier
- cell death
- skeletal muscle
- signaling pathway
- long non coding rna
- wild type
- cell proliferation