Examination of Endogenous Rotund Expression and Function in Developing Drosophila Olfactory System Using CRISPR-Cas9-Mediated Protein Tagging.
Abstract
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.
Type
Journal articleSubject
CRISPRCas9
genome editing
homologous recombination
olfactory system development
rotund
tagging
Animals
CRISPR-Cas Systems
Drosophila
Drosophila Proteins
Gene Expression
Gene Expression Regulation, Developmental
Gene Targeting
Homologous Recombination
Olfactory Bulb
Protein Transport
RNA Editing
RNA, Guide
Transcription Factors
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https://hdl.handle.net/10161/10914Published Version (Please cite this version)
10.1534/g3.115.021857Publication Info
Li, 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.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Pelin Cayirlioglu Volkan
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

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