1. Developmental Biology
  2. Neuroscience
Download icon

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
Tools and Resources
  • Cited 10
  • Views 3,874
  • Annotations
Cite this article as: eLife 2017;6:e26420 doi: 10.7554/eLife.26420

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

Publication 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.

Metrics

  • 3,874
    Page views
  • 664
    Downloads
  • 10
    Citations

Article citation count generated by polling the highest count across the following sources: PubMed Central, Crossref, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Developmental Biology
    2. Neuroscience
    Ryan T Willett et al.
    Research Article Updated
    1. Developmental Biology
    2. Plant Biology
    Carla Verna et al.
    Research Article