Conditional knockout of retinal determination genes in differentiating cells in Drosophila
Meng Jina b, Aiden Eblimitc d e f, Merlyn X Pulikkatharag h, Stuart J. Corrg h i j k l, Rui Chenc d e f m n, Graeme Mardona b c e m n o p q r s✉
- aDepartment of Pathology and Immunology Baylor College of Medicine Houston TX USA
- bDepartment of Pathology and Immunology, Baylor College of Medicine, Houston, TX, USA
- cDepartment of Molecular and Human Genetics Baylor College of Medicine Houston TX USA
- dHuman Genome Sequencing Center Baylor College of Medicine Houston TX USA
- eDepartment of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA
- fHuman Genome Sequencing Center, Baylor College of Medicine, Houston, TX, USA
- gDepartment of Surgery Baylor College of Medicine Houston TX USA
- hDepartment of Surgery, Baylor College of Medicine, Houston, TX, USA
- iDepartment of Biomedical Engineering University of Houston TX USA
- jDepartment of Chemistry Rice University Houston TX USA
- kDepartment of Biomedical Engineering, University of Houston, TX, USA
- l
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Abstract
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.