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Elife. 2017 May 31;6. pii: e26420. doi: 10.7554/eLife.26420.

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

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Howard Hughes Medical Institute, Baylor College of Medicine, Houston, United States.
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, United States.
Department of Neuroscience, Baylor College of Medicine, Houston, United States.
Program in Developmental Biology, Baylor College of Medicine, Houston, United States.
Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, United States.


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, 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.


D. melanogaster; FLEx; MiMIC; SNF4Aγ; Trim9; developmental biology; effete; neuroscience; post-mitotic cells; stem cells

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