Examination of Endogenous Rotund Expression and Function in Developing Drosophila Olfactory System Using CRISPR-Cas9-Mediated Protein Tagging.
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The zinc-finger protein Rotund (Rn) plays a critical role in controlling the development of the fly olfactory system. However, little is known about its molecular function in vivo. Here, we added protein tags to the rn locus using CRISPR-Cas9 technology in Drosophila to investigate its subcellular localization and the genes that it regulates . We previously used a reporter construct to show that rn is expressed in a subset of olfactory receptor neuron (ORN) precursors and it is required for the diversification of ORN fates. Here, we show that tagged endogenous Rn protein is functional based on the analysis of ORN phenotypes. Using this method, we also mapped the expression pattern of the endogenous isoform-specific tags in vivo with increased precision. Comparison of the Rn expression pattern from this study with previously published results using GAL4 reporters showed that Rn is mainly present in early steps in antennal disc patterning, but not in pupal stages when ORNs are born. Finally, using chromatin immunoprecipitation, we showed a direct binding of Rotund to a previously identified regulatory element upstream of the bric-a-brac gene locus in the developing antennal disc.
olfactory system development
Gene Expression Regulation, Developmental
Published Version (Please cite this version)10.1534/g3.115.021857
Publication InfoLi, Qingyun; Barish, Scott; Okuwa, Sumie; & Volkan, Pelin C (2015). Examination of Endogenous Rotund Expression and Function in Developing Drosophila Olfactory System Using CRISPR-Cas9-Mediated Protein Tagging. G3 (Bethesda), 5(12). pp. 2809-2816. 10.1534/g3.115.021857. Retrieved from https://hdl.handle.net/10161/10914.
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Associate Professor of Biology
The long-term goal in the lab is to understand the developmental processes that establish the basic organizational and functional principles of the neuronal circuits in the brain. We investigate how the neuronal circuits assemble, functionally mature, remodel in developmental and evolutionary time scales. To understand these processes the Volkan lab uses the olfactory system of the genetically tractable Drosophila melanogaster as a model organism and apply molecular, developmental and systems le