12:20pm
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1:20pm |
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VTLSS Seminar: Relief of Period2 post-transcriptional regulation by 3-UTR
(Research)
Location:
Biocomplexity Institute of Virginia Tech, Conference Center (145)
Presenter:
Seung-Hee Yoo
Assistant Professor, University of Texas
Host:
Shihoko Kojima
Fellow and Assistant Professor, Virginia Tech
Abstract: We previously reported a circadian reporter knockin mouse line, Per2::Luc, which expresses PER2::LUC fusion proteins and has been widely used for real-time circadian bioluminescence monitoring. To understand in vivo function of Per2 3'-UTR, we generated Per2::LucSV, a modified knockin reporter mouse line where the endogenous Per2 3'-UTR was replaced by an SV40 late poly(A) signal.
Bioluminescence recording and molecular analysis showed robust PER2::LUC rhythms in various tissues with significantly enhanced amplitude compared with those in Per2::Luc. In addition, Per2::LucSV mice also exhibited lengthened free-running periods (~24.0 hr), greater phase delays following light pulse, and enhanced temperature compensation relative to Per2::Luc. Analysis of the Per2 3'-UTR sequence revealed that miR-24, and to a lesser degree miR-30, suppressed PER2 protein translation, and the relief of this inhibition in Per2::LucSV augmented PER2::LUC protein level and oscillatory amplitude.
Together, these studies provide important mechanistic insights into the regulatory roles of miR-24 and PER2 in Per2 expression and core clock function, and establish Per2::LucSV as a valuable in vivo system endowed with enhanced bioluminescence and circadian amplitude.
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