Abstract
The fruit fly, Drosophila melanogaster, has been a favorite experimental system of developmental biologists for more than a century. One of the most attractive features of this model system is the clarity by which one can analyze mutant phenotypes. Most genes are found in single copies, and loss-of-function mutants often have obvious phenotypes that can be analyzed during development and in adulthood. As with all metazoans, a significant fraction of Drosophila genes are used during both embryonic and postembryonic development, and null mutants often die during embryogenesis thereby precluding the analysis of postembryonic tissues. For several decades researchers worked around this problem by either studying gynandromorphs or irradiating chromosomes carrying mutations in the hope of inducing mitotic recombination which would then allow for the analysis of mutant phenotypes in smaller populations of cells. The former method suffers from the fact that mutations in the gene of interest are often lethal when generated in large sectors, which is a hallmark of gynandromorphs. Clonal induction with the latter method occurs at relatively low frequencies making this method laborious. The introduction of the yeast FRT System/FRT site-directed recombination system to Drosophila has made generating loss-of-function mosaic clones simple and easy. Over the years several variants of this method have allowed developmental biologists to remove genes, overexpress genes, and even express one gene in patches of cells that are mutant for a second gene. In this review we will briefly discuss some of various FRT System/FRT-based approaches that are being used to manipulate gene expression in Drosophila. The individual FRT System/FRT-based methods are described in the papers that are cited herein. We will outline the procedure that our lab uses to prepare and analyze mosaic clones in Drosophila eye-antennal imaginal discs.
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Acknowledgments
We would like to thank Donald Ready and Kevin Moses for the original protocol for the dissection of eye-antennal imaginal discs. This work is supported by a grant from the National Eye Institute (R01 EY014863) to Justin P. Kumar.
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Weasner, B.M., Zhu, J., Kumar, J.P. (2017). FLPing Genes On and Off in Drosophila . In: Eroshenko, N. (eds) Site-Specific Recombinases. Methods in Molecular Biology, vol 1642. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7169-5_13
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DOI: https://doi.org/10.1007/978-1-4939-7169-5_13
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