Date of Completion
iPSC, Stem Cell, CRISPR/Cas9, Angelman Syndrome, Imprinting, Long non-coding RNA, Prader-Willi Syndrome, Chromatin structure
Field of Study
Doctor of Philosophy
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.
Hsiao, Jack Shuo-Hung, "Understanding the Regulation of UBE3A-ATS and the Process of UBE3A Imprinting Using iPSC Modeling and CRISPR/Cas9 Editing" (2016). Doctoral Dissertations. 1274.