Show simple item record

dc.contributor.advisorWolfe, Michael S.
dc.contributor.authorDickson, John Robert
dc.date.accessioned2013-10-18T13:56:59Z
dash.embargo.terms2015-10-10en_US
dash.embargo.terms2015-10-10
dc.date.issued2013-10-18
dc.date.submitted2013
dc.identifier.citationDickson, John Robert. 2013. The Role of the Human Tau 3'-Untranslated Region in Regulating Tau Expression. Doctoral dissertation, Harvard University.en_US
dc.identifier.otherhttp://dissertations.umi.com/gsas.harvard:11070en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:11181167
dc.description.abstractThe microtubule-associated protein tau forms pathological neuronal filaments in Alzheimer's disease (AD) and other neurodegenerative disorders, known collectively as tauopathies. Previous studies in transgenic mouse models of AD suggest that reducing tau expression may be safe and beneficial for the prevention or treatment of AD and possibly other tauopathies. As a first step toward identifying novel therapeutic strategies to reduce tau levels, the studies presented in this dissertation aim to investigate the role of the human tau 3'-untranslated region (3'-UTR) in regulating tau expression. Tau expresses two 3'-UTR isoforms, long and short, as a result of alternative polyadenylation. The exact sequence of these two 3'-UTR isoforms was determined by rapid amplification of cDNA 3'-ends (3'-RACE), and the two 3'-UTR isoforms were cloned into a luciferase reporter vector. Using these reporter constructs, the expression of these isoforms was found to be differentially controlled in human neuroblastoma cell lines M17D and SH-SY5Y by luciferase assays and quantitative PCR (qPCR). Through an unbiased screen of tau 3'-UTR deletions and fragments using luciferase reporter constructs, several regions in the long tau 3'-UTR isoform that contain regulatory cis-elements were identified. Additionally, several microRNAs were computationally identified as candidates that might bind the long tau 3'-UTR and thereby differentially control the expression of long versus short tau 3'-UTR isoforms. Screening these candidate microRNAs via luciferase reporter assay identified miR-34a, which was subsequently shown to repress the expression of endogenous tau protein and mRNA in M17D cells using Western blot and qPCR, respectively. Conversely, inhibition of endogenously expressed miR-34 family members leads to increased endogenous tau expression. Taken together, these studies suggest that the expression of the two tau 3'-UTR isoforms is differentially regulated and that this differential regulation is due to the presence of regulatory cis-elements found only in the long tau 3'-UTR isoform, including a binding site for miR-34 family members. Improved understanding of the regulation of tau expression by its 3'-UTR may ultimately lead to the development of novel therapeutic strategies for the treatment of Alzheimer's disease and other tauopathies.en_US
dc.language.isoen_USen_US
dash.licenseLAA
dc.subjectMolecular biologyen_US
dc.subjectNeurosciencesen_US
dc.subject3'-untranslated regionen_US
dc.subjectAlternative polyadenylationen_US
dc.subjectAlzheimer's diseaseen_US
dc.subjectMAPTen_US
dc.subjectmiR-34aen_US
dc.subjectTauen_US
dc.titleThe Role of the Human Tau 3'-Untranslated Region in Regulating Tau Expressionen_US
dc.typeThesis or Dissertationen_US
dash.depositing.authorDickson, John Robert
dc.date.available2015-10-10T07:33:04Z
thesis.degree.date2013en_US
thesis.degree.disciplineBiology: Medical Sciences, Division ofen_US
thesis.degree.grantorHarvard Universityen_US
thesis.degree.leveldoctoralen_US
thesis.degree.namePh.D.en_US
dc.contributor.committeeMemberBuratowski, Stephenen_US
dc.contributor.committeeMemberGregory, Richarden_US
dc.contributor.committeeMemberKrichevsky, Annaen_US
dc.contributor.committeeMemberTsai, Li-Hueien_US
dash.contributor.affiliatedDickson, John


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record