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.
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Li, Qingyun, Scott Barish, Sumie Okuwa and Pelin C Volkan (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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Pelin Cayirlioglu Volkan
The primary intellectual focus of our lab centers on unraveling the molecular and circuit mechanisms through which social experiences mold the brains and responses of organisms. To investigate these phenomena, we employ the fruit fly nervous system as a model and take an interdisciplinary approach that integrates genetic, behavioral, circuit-mapping, and systems-level molecular tools. Recent advancements in neurogenetics and neuro-visualization techniques in Drosophila melanogaster, a model system with a rich history in behavioral and neurogenetic research, provide us with unique and unprecedented advantages for exploring these questions. Within the realm of fruit flies, several noteworthy observations emerge: 1) Social isolation exerts significant effects on the Drosophila brain and behaviors, 2) well-established connections exist between genes, neural circuits, and stereotyped social behaviors, 3) the utilization of gene editing and neuronal circuit mapping methods is unparalleled, and 4) these resources are further enriched by existing and upcoming connectome data. Leveraging this comprehensive toolset, our overarching objective is to identify genes regulated by social isolation, determine their expression and function in individual neurons and circuits in the brain, and ascertain how variations in these processes influence both brain function and behavioral responses to isolation.
Questions we are interested in:
1- How does social experience and pheromone circuit activity modulate gene expression in the nervous system?
2- How does social experience and pheromone circuit activity modulate circuit structure and function?
3- How does social experience and pheromone circuit activity modulate behaviors like locomotion, feeding, courtship and aggression?
4- How does social experience and pheromone circuit activity modulate physiology like metabolism, circulatory system and immunity?
5- What makes individuals more sensitive or resilient to the effects of social experience?
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