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.
- Sonal Nagarkar-Jaiswal
- Hugo J Bellen
- Sathiya N Manivannan
- Zhongyuan Zuo
- Zhongyuan Zuo
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
- Mani Ramaswami, Trinity College Dublin, Ireland
© 2017, Nagarkar-Jaiswal et al.
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