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

Abstract
Article and author information
Metrics

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

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.
Sathiya N Manivannan

Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States

Competing interests
No competing interests declared.
Zhongyuan Zuo

Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States

Competing interests
No competing interests declared.
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.

"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.

Copyright

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.