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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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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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.
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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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