Functional characterization of Mest and its imprinted gametic differentially methylated region
department or school: Medicine, Faculty of
department or school: Medical Genetics, Department of
data provider: DSpace
degree campus: UBCV
creator: Ha, Amanda
issued: 2022-12-12T19:31:52Z
issued: 2022
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: Genomic imprinting is an epigenetic phenomenon by which the expression of certain genes follows a parent-of-origin-specific pattern. The sub-proximal end of mouse Chr 6 contains an imprinted domain comprised of the paternally expressed gene Mest, and the maternally expressed genes Copg2 and Klf14. The Mest locus harbors the only gametic differentially methylated region (gDMR) in the region, which may act as an Imprinting Control Region for the coordinated regulation of these three imprinted genes. The Mest-Klf14 imprinted domain shares syntenic homology with human Chr 7q32. Therefore, loss of Mest function, or Klf14 overexpression may contribute to the maternal uniparental disomy 7 (mUPD7) growth retardation phenotype associated with Sliver- Russell syndrome (SRS). Here, I investigate the function of Mest, and its gDMR. In Chapter 3, I leveraged a previously generated Mest KO mouse line to characterize the function of Mest in developmental pathways. Embryonic growth retardation observed in Mest KO mice was independent of gross placental abnormalities in vivo, but associated with altered Wnt signalling in cell assays. Based on additional proteomic and metabolomic characterization, I hypothesize that the putative epoxide hydrolase, known as MEST, acts upon metabolites within the endoplasmic reticulum lumen. In Chapter 4, I explored the function of the Mest gDMR. While the imprinted regulation of both Mest and Copg2 has been well documented, the mechanism of Klf14 imprinting is unknown. Klf14 expression is lost in embryos lacking maternal methylation iii genome-wide, indicating that maternally-inherited DNA methylation (DNAme) is paradoxically required for its expression. Through CRISPR-Cas9 mutagenesis and allele-specific chromosome conformation experiments in F1 embryonic stem cells, I demonstrate that the Mest gDMR regulates imprinting of Klf14 through long-range chromatin looping, permitting differential promoter-enhancer interactions between parental alleles. I pr…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/83395
note: FUNCTIONAL CHARACTERIZATION OF MEST AND ITS IMPRINTED GAMETIC DIFFERENTIALLY METHYLATED REGION by Amanda Ha B.Sc., The University of Victoria, 2015 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Medical Genetics) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) December 2022 © Amanda Ha, 2022 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the dissertation entitled: Functional characterization of Mest and its imprinted gametic differentially methylated region submitted by Amanda Ha in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Medical Genetics Examining Committee: Dr. Louis Lefebvre, Associate Professor, Medical Genetics, UBC Supervisor Dr. Carolyn Brown, Professor, Medical Genetics, UBC Supervisory Committee Member Dr. LeAnn Howe, Professor, Biochemistry and Molecular Biology, UBC University Examiner Dr. Wendy Robinson, Professor, Medical Genetics, UBC University Examiner Dr. Marisa Bartolomei, Professor, Cell and Developmental Biology, University of Pennsylvania External Examiner Additional Supervisory Committee Members: Dr. T. Michael Underhill, Professor, Cellular and Physiological Science, UBC Supervisory Committee Member Dr. Pamela Hoodless, Professor, Medical Genetics, UBC Supervisory Committee Member iii Abstract Genomic imprinting is an epigenetic phenomenon by which the expression of certain genes follows a parent-of-origin-specific pattern. The sub-proximal end of mouse Chr 6 contains an imprinted domain comprised of the paternally expressed gene Mest, and the maternally expressed genes Copg2 and Klf14. The Mest locus harbors the only gametic differentially methylated region (gDMR) in the region, which may act as an Imprinting Control Region for the coordinated regulation of these three imprinted genes. The Mest-Klf14 impri…
has type: Thesis/Dissertation
date issued: 2023-05
is shown at: 10.14288/1.0422487
language: eng
degree discipline: Medical Genetics
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Lefebvre, Louis
type: Text
identifier URI: http://hdl.handle.net/2429/83395