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The twin spot generator for differential Drosophila lineage analysis

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Abstract

In Drosophila melanogaster, widely used mitotic recombination–based strategies generate mosaic flies with positive readout for only one daughter cell after division. To differentially label both daughter cells, we developed the twin spot generator (TSG) technique, which through mitotic recombination generates green and red twin spots that are detectable after the first cell division as single cells. We propose wide applications of TSG to lineage and genetic mosaic studies.

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Figure 1: TSG strategy.
Figure 2: Red and green twin spots, and yellow clones, generated after mitotic recombination at 82F7.
Figure 3: Separation of clones in developing leg imaginal discs.

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Acknowledgements

We thank J. Zirin and M. Packard for confocal expertise, P. Bradley for molecular biology advice, F. Karch (University of Geneva) for providing flies carrying integrase and T.S. Griffin for contributing the technique acronym. This work was supported by grants from the US National Institutes of Health (1 RO1 GM61936, C.-t.W.; RO1 GM058282, G.S.; and RO1 GM084947, N.P.) and a Ruth L. Kirschstein National Research Service award (1 F32 GM67460, J.R.B.); the US National Science Foundation (A.M.H.); the Swiss National Science Foundation (PBBE33-121069, M.B.) and the Leukemia and Lymphoma Society (C.B.).

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Author notes

  1. Amber M Hohl & Jack R Bateman

    Present address: Present addresses: Department of Biochemistry, University of Iowa, Iowa City, Iowa, USA (A.M.H.) and Biology Department, Bowdoin College, Brunswick, Maine, USA (J.R.B.).,

Authors and Affiliations

  1. Commissariat à l'Energie Atomique, Direction des Sciences du Vivant, Institut de Recherches en Technologies et Sciences pour le Vivant, Laboratoire Biochimie et Biophysique des Systèmes Intégrés, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5092, and Université Joseph Fourier, Grenoble, France

    Ruth Griffin

  2. Department of Biology, University of Washington, Seattle, Washington, USA

    Anne Sustar, Marianne Bonvin & Gerold Schubiger

  3. Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA

    Richard Binari, Amber M Hohl, Jack R Bateman, Christians Villalta, Elleard Heffern, Chris Bakal, C-ting Wu & Norbert Perrimon

  4. Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts, USA

    Richard Binari, Christians Villalta, Elleard Heffern, Chris Bakal & Norbert Perrimon

  5. Department of Biology, Center for Developmental Genetics, New York University, New York, New York, USA

    Alberto del Valle Rodriguez & Claude Desplan

  6. Laboratoire Transduction de Signal, Unité 873, Institut National de la Santé et de la Recherche Médicale, Commissariat à l'Energie Atomique, Direction des Sciences du Vivant, Institut de Recherches en Technologies et Sciences pour le Vivant, Grenoble, France

    Didier Grunwald

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  1. Ruth Griffin
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  8. Christians Villalta
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Contributions

R.G., design and execution of constructs, vectors and first-round proof-of-principle experiments, verification of TSG lines and writing of manuscipt. A.S., M.B. and G.S., clonal separation experiments and description in manuscript. R.B., design and generation of TSG transgenic fly lines and modification of target genomic lines. A.d.V.R., validation of TSG in fly brain. C.B., molecular biology expertise and tissue culture strategy. A.M.H., validation of TSG in imaginal discs. J.R.B., design and generation of target genomic lines. C.V., fly injections. E.H., tissue culture assays. D.G., acquisition and analysis of confocal images. C.D., G.S., C.-t.W. and N.P. co-directed the project. C.-t.W. suggested and funded the RMCE approach. N.P. provided US laboratory facilities and materials and conceived the TSG strategy.

Corresponding authors

Correspondence to Ruth Griffin, C-ting Wu or Norbert Perrimon.

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Supplementary Figures 1–4, Supplementary Tables 1–5 and Supplementary Note (PDF 3410 kb)

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Griffin, R., Sustar, A., Bonvin, M. et al. The twin spot generator for differential Drosophila lineage analysis. Nat Methods 6, 600–602 (2009). https://doi.org/10.1038/nmeth.1349

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  • DOI: https://doi.org/10.1038/nmeth.1349

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