A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells

  1. Sonal Nagarkar-Jaiswal  Is a corresponding author
  2. Sathiya N Manivannan
  3. Zhongyuan Zuo
  4. Hugo J Bellen  Is a corresponding author
  1. Howard Hughes Medical Institute, Baylor College of Medicine, United States
  2. Baylor College of Medicine, United States

Abstract

Here we describe a novel method based on intronic MiMIC insertions described in Nagarkar-Jaiswal et al. (2015) to perform conditional gene inactivation in Drosophila. Mosaic analysis in Drosophila cannot be easily performed in post-mitotic cells. We therefore developed Flip-Flop, a flippase-dependent in vivo cassette-inversion method that marks wild-type cells with the endogenous EGFP-tagged protein, whereas mutant cells are marked with mCherry upon inversion. We document the ease and usefulness of this strategy in differential tagging of wild-type and mutant cells in mosaics. We use this approach to phenotypically characterize the loss of SNF4Aγ, encoding the γ subunit of the AMP Kinase complex. The Flip-Flop method is efficient and reliable, and permits conditional gene inactivation based on both spatial and temporal cues, in a cell cycle-, and developmental stage-independent fashion, creating a platform for systematic screens of gene function in developing and adult flies with unprecedented detail.

Article and author information

Author details

  1. Sonal Nagarkar-Jaiswal

    Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
    For correspondence
    snagarka@bcm.edu
    Competing interests
    No competing interests declared.
  2. Sathiya N Manivannan

    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
    Competing interests
    No competing interests declared.
  3. Zhongyuan Zuo

    Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States
    Competing interests
    No competing interests declared.
  4. Hugo J Bellen

    Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States
    For correspondence
    hbellen@bcm.edu
    Competing interests
    Hugo J Bellen, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5992-5989

Funding

Howard Hughes Medical Institute

  • Sonal Nagarkar-Jaiswal
  • Hugo J Bellen

National Institute of Neurological Disorders and Stroke (NINDS U54 NS093793)

  • Sathiya N Manivannan

Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD U54 HD083082)

  • Zhongyuan Zuo

Robert A. and Renee E. Belfer Family Foundation

  • Zhongyuan Zuo

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

Reviewing Editor

  1. Mani Ramaswami, Trinity College Dublin, Ireland

Version history

  1. Received: February 28, 2017
  2. Accepted: May 25, 2017
  3. Accepted Manuscript published: May 31, 2017 (version 1)
  4. Version of Record published: June 30, 2017 (version 2)

Copyright

© 2017, Nagarkar-Jaiswal 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. Sonal Nagarkar-Jaiswal
  2. Sathiya N Manivannan
  3. Zhongyuan Zuo
  4. Hugo J Bellen
(2017)
A cell cycle-independent, conditional gene inactivation strategy for differentially tagging wild-type and mutant cells
eLife 6:e26420.
https://doi.org/10.7554/eLife.26420

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https://doi.org/10.7554/eLife.26420

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