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

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.

Data availability

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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  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
(2021)
tTARGIT AAVs mediate the sensitive and flexible manipulation of intersectional neuronal populations in mice
eLife 10:e66835.
https://doi.org/10.7554/eLife.66835

Further reading

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    Using screen counts, women 50–64 years old have lower cancer screening rates for cervical and colorectal cancers (CRC) than all other age ranges. This paper aims to present woman-centric cervical cancer and CRC screenings to determine the predictor of being up-to-date for both.

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    We used the Behavioral Risk Factor Surveillance System (BRFSS), an annual survey to guide health policy in the United States, to explore the up-to-date status of dual cervical cancer and CRC screening for women 50–64 years old. We categorized women into four mutually exclusive categories: up-to-date for dual-screening, each single screen, or neither screen. We used multinomial multivariate regression modeling to evaluate the predictors of each category.

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    Among women ages 50–64 years old, dual-screening was reported for 58.2% (57.1–59.4), cervical cancer screening alone (27.1% (26.0–28.2)), CRC screening alone (5.4% (4.9–5.9)), and neither screen (9.3% (8.7–9.9)). Age, race, education, income, and chronic health conditions were significantly associated with dual-screening compared to neither screen. Hispanic women compared to non-Hispanic White women were more likely to be up-to-date with cervical cancer screening than dual-screening (adjusted odds ratio [aOR] = 1.39 (1.10, 1.77)). Compared to younger women, those 60–64 years are significantly more likely to be up-to-date with CRC screening than dual-screening (aOR = 1.75 (1.30, 2.35)).

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    Screening received by each woman shows a much lower rate of dual-screening than prior single cancer screening rates. Addressing dual-screening strategies rather than single cancer screening programs for women 50–64 years may increase both cancer screening rates.

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    This work was supported by NIH through the Michigan Institute for Clinical and61 Health Research UL1TR002240 and by NCI through The University of Michigan Rogel Cancer62 Center P30CA046592 grants.

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    Supported by National Institutes of Health R01 HL122352 grant; ‘la Caixa’ Banking Foundation (HR18-00304); Fundación La Marató TV3: Ayudas a la investigación en enfermedades raras 2020 (LA MARATO-2020); Instituto de Salud Carlos III/FEDER/FSE; Horizon 2020 - Research and Innovation Framework Programme GA-965286 to JJ; the CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the Ministerio de Ciencia e Innovación (MCIN) and the Pro CNIC Foundation), and is a Severo Ochoa Center of Excellence (grant CEX2020-001041-S funded by MICIN/AEI/10.13039/501100011033). American Heart Association postdoctoral fellowship 19POST34380706s to JVEN. Israel Science Foundation to OB and MA [824/19]. Rappaport grant [01012020RI]; and Niedersachsen Foundation [ZN3452] to OB; US-Israel Binational Science Foundation (BSF) to OB and TH [2019039]; Dr. Bernard Lublin Donation to OB; and The Duchenne Parent Project Netherlands (DPPNL 2029771) to OB. National Institutes of Health R01 AR068428 to DM and US-Israel Binational Science Foundation Grant [2013032] to DM and OB.