Date of Completion

9-21-2016

Embargo Period

3-15-2017

Keywords

iPSC, Stem Cell, CRISPR/Cas9, Angelman Syndrome, Imprinting, Long non-coding RNA, Prader-Willi Syndrome, Chromatin structure

Major Advisor

Stormy Chamberlain

Associate Advisor

Marc Lalande

Associate Advisor

Gordon Carmichael

Associate Advisor

Brenton Graveley

Associate Advisor

Rogina Blanka

Field of Study

Biomedical Science

Degree

Doctor of Philosophy

Open Access

Open Access

Abstract

Angelman Syndrome (AS) is a neurodevelopmental disorder caused by loss of the maternal copy of UBE3A. The paternal copy cannot compensate for the loss because it is subject to tissue-specific imprinting in the brain. This imprinting is controlled by the reciprocal UBE3A antisense transcript (UBE3A-ATS) expressed only in the brain. The goal of my thesis projects is to understand the underlying mechanism by which the UBE3A-ATS is regulated. We found that UBE3A-ATS is expressed and UBE3A is imprinted in non-neurons from an individual with 187 kb deletion at the paternal allele. This suggests that expression of UBE3A-ATS does not require any neuronal factors, and the regulatory elements reside in the genomic region. A minimal region consisted of the bipartite boundary element IPW and PWAR1 is identified using CRISPR/Cas9. Absence of this region leads to higher UBE3A-ATS expression and early UBE3A imprinting during neural differentiation. SNRPN-PWAR1-UBE3A is in close proximity in 3D, but this interaction largely remains the same during neural differentiation. My thesis work not only demonstrates how UBE3A-ATS is regulated, but also provides evidence that interfering with UBE3A-ATS transcription reactivates UBE3A.

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