Conditional knockout of retinal determination genes in differentiating cells in Drosophila.

TitleConditional knockout of retinal determination genes in differentiating cells in Drosophila.
Publication TypeJournal Article
Year of Publication2016
AuthorsJin, M, Eblimit, A, Pulikkathara, M, Corr, S, Chen, R, Mardon, G
JournalFEBS J
Date Published2016 Aug
KeywordsAlleles, Animals, Cell Differentiation, Drosophila, Drosophila Proteins, Eye, Eye Proteins, Gene Knockout Techniques, Homeodomain Proteins, Larva, Photoreceptor Cells, Invertebrate, Retina

Conditional gene knockout in postmitotic cells is a valuable technique which allows the study of gene function with spatiotemporal control. Surprisingly, in contrast to its long-term and extensive use in mouse studies, this technology is lacking in Drosophila. Here, we use a novel method for generating complete loss of eyes absent (eya) or sine oculis (so) function in postmitotic cells posterior to the morphogenetic furrow (MF). Specifically, genomic rescue constructs with flippase recognition target (FRT) sequences flanking essential exons are used to generate conditional null alleles. By removing gene function in differentiating cells, we show that eya and so are dispensable for larval photoreceptor differentiation, but are required for differentiation during pupal development. Both eya and so are necessary for photoreceptor survival and the apoptosis caused by loss of eya or so function is likely a secondary consequence of inappropriate differentiation. We also confirm their requirement for cone cell development and reveal a novel role in interommatidial bristle (IOB) formation. In addition, so is required for normal eye disc morphology. This is the first report of a knockout method to study eya and so function in postmitotic cells. This technology will open the door to a large array of new functional studies in virtually any tissue and at any stage of development or in adults.

Alternate JournalFEBS J
PubMed ID27257739
PubMed Central IDPMC4975629
Grant ListP30 EY002520 / EY / NEI NIH HHS / United States
R01 EY011232 / EY / NEI NIH HHS / United States

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