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Examination of Endogenous Rotund Expression and Function in Developing Drosophila Olfactory System Using CRISPR-Cas9-Mediated Protein Tagging.

dc.contributor.author Li, Q
dc.contributor.author Barish, S
dc.contributor.author Okuwa, S
dc.contributor.author Volkan, PC
dc.coverage.spatial United States
dc.date.accessioned 2015-11-19T17:37:12Z
dc.date.issued 2015-10-23
dc.identifier http://www.ncbi.nlm.nih.gov/pubmed/26497147
dc.identifier g3.115.021857
dc.identifier.uri http://hdl.handle.net/10161/10914
dc.description.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.
dc.language eng
dc.relation.ispartof G3 (Bethesda)
dc.relation.isversionof 10.1534/g3.115.021857
dc.subject CRISPR
dc.subject Cas9
dc.subject genome editing
dc.subject homologous recombination
dc.subject olfactory system development
dc.subject rotund
dc.subject tagging
dc.subject Animals
dc.subject CRISPR-Cas Systems
dc.subject Drosophila
dc.subject Drosophila Proteins
dc.subject Gene Expression
dc.subject Gene Expression Regulation, Developmental
dc.subject Gene Targeting
dc.subject Homologous Recombination
dc.subject Olfactory Bulb
dc.subject Protein Transport
dc.subject RNA Editing
dc.subject RNA, Guide
dc.subject Transcription Factors
dc.title Examination of Endogenous Rotund Expression and Function in Developing Drosophila Olfactory System Using CRISPR-Cas9-Mediated Protein Tagging.
dc.type Journal article
pubs.author-url http://www.ncbi.nlm.nih.gov/pubmed/26497147
pubs.begin-page 2809
pubs.end-page 2816
pubs.issue 12
pubs.organisational-group Basic Science Departments
pubs.organisational-group Biology
pubs.organisational-group Duke
pubs.organisational-group Duke Institute for Brain Sciences
pubs.organisational-group Institutes and Provost's Academic Units
pubs.organisational-group Neurobiology
pubs.organisational-group School of Medicine
pubs.organisational-group Trinity College of Arts & Sciences
pubs.organisational-group University Institutes and Centers
pubs.publication-status Published online
pubs.volume 5
dc.identifier.eissn 2160-1836


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