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

  1. Paul V Sabatini
  2. Jine Wang
  3. Alan C Rupp
  4. Alison H Affinati
  5. Jonathan N Flak
  6. Chien Li
  7. David P Olson
  8. Martin G Myers Jr  Is a corresponding author
  1. University of Michigan, United States
  2. University of Indiana, United States
  3. Novo Nordisk Research Center, United States
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Cite this article as: eLife 2021;10:e66835 doi: 10.7554/eLife.66835

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 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.

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

Article and author information

Author details

  1. Paul V Sabatini

    Internal Medicine, University of Michigan, Ann Arbor, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6613-566X
  2. Jine Wang

    Internal Medicine, University of Michigan, Ann Arbor, United States
    Competing interests
    No competing interests declared.
  3. Alan C Rupp

    Internal Medicine, University of Michigan, Ann Arbor, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5363-4494
  4. Alison H Affinati

    Internal Medicine, University of Michigan, Ann Arbor, United States
    Competing interests
    No competing interests declared.
  5. Jonathan N Flak

    Pharmacology and Toxicology, University of Indiana, Indianapolis, United States
    Competing interests
    No competing interests declared.
  6. Chien Li

    Obesity, Novo Nordisk Research Center, Seattle, United States
    Competing interests
    Chien Li, CL is an employee of Novo Nordisk A/S..
  7. David P Olson

    Internal Medicine, University of Michigan, Ann Arbor, United States
    Competing interests
    No competing interests declared.
  8. 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.
    ORCID icon "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.

Reviewing Editor

  1. Richard D Palmiter, Howard Hughes Medical Institute, University of Washington, United States

Publication history

  1. Received: January 23, 2021
  2. Accepted: March 10, 2021
  3. Accepted Manuscript published: March 11, 2021 (version 1)
  4. Version of Record published: April 7, 2021 (version 2)

Copyright

© 2021, Sabatini et al.

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.

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    Funding: This work was supported in part by the Italian Ministry of Health (GR-2013-02355724 and Ricerca Corrente), MJFF, ALZ, Alzheimer's Research UK and the Weston Brain Institute (BAND2015), and Euronanomed III (Speedy) to FM; by the Spanish Ministerio de Economía y Competitividad [grant AGL2016-78054-R (AEI/FEDER, UE)] to J.M.T. and J.C.E.; A.M.-M. was supported by a fellowship from the INIA (FPI-SGIT-2015-02).

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