tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice

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Abstract

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) AAVs. tTARGIT AAVs utilize a Flp-dependent tetracycline transactivator (tTA) 'Driver' AAV and a tetracycline response element (TRE)-driven, Cre-dependent 'Payload' AAV to express the transgene of interest. We employed this system in Slc17a6^FlpO;Lepr^Cre mice to manipulate LepRb neurons of the ventromedial hypothalamus (VMH; LepRb^VMH neurons) while omitting neighboring LepRb populations. We defined the circuitry of LepRb^VMH 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.

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All data generated or analysed during this study are included in the manuscript and supporting files

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Paul V Sabatini

Internal Medicine, University of Michigan, Ann Arbor, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-6613-566X
Jine Wang

Internal Medicine, University of Michigan, Ann Arbor, United States

Competing interests
No competing interests declared.
Alan C Rupp

Internal Medicine, University of Michigan, Ann Arbor, United States

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-5363-4494
Alison H Affinati

Internal Medicine, University of Michigan, Ann Arbor, United States

Competing interests
No competing interests declared.
Jonathan N Flak

Pharmacology and Toxicology, University of Indiana, Indianapolis, United States

Competing interests
No competing interests declared.
Chien Li

Obesity, Novo Nordisk Research Center, Seattle, United States

Competing interests
Chien Li, CL is an employee of Novo Nordisk A/S..
David P Olson

Internal Medicine, University of Michigan, Ann Arbor, United States

Competing interests
No competing interests declared.
Martin G Myers Jr

Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, United States

For correspondence
mgmyers@umich.edu

Competing interests
No competing interests declared.

"This ORCID iD identifies the author of this article:" 0000-0001-9468-2046

Funding

American Diabetes Association (1-19-PDF-099)

Paul V Sabatini

China scholarship council (201908420207)

Jine Wang

National Institute of Diabetes and Digestive and Kidney Diseases (DK104999)

David P Olson

National Institute of Diabetes and Digestive and Kidney Diseases (DK056731)

Martin G Myers Jr

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

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This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.