Regulation of UBE3A Imprinted Expression

摘要 UBE3A的印记表达是神经元特异性的，发生是因为父系遗传的等位基因是 沉默了。一个长的非编码反义转录本UBE3A-ATS以神经元特异的方式表达，并 通过一种未知的机制介导UBE3A印迹。这项建议的总体目标是理解 UBE3A-ATS神经元特异性表达调控和抑制的过程 父系UBE3A在人类神经元中的表达。我们建议研究三个不同的方面来规范UBE3A 印迹表达：1)人类神经元中编码与非编码Snrpn RNA的调节；2) 对UBE3A-ATS，Snrpn ncRNA的末端部分的组织特异性调节；以及3)通过 哪个UBE3A-ATS导致父本UBE3A的抑制。我们将使用患者特定的诱导多能性 Angelman综合征(AS)患者来源的干细胞(IPSCs)及其神经元衍生品 学习。我们将剖析制约UBE3A-ATS的顺式作用边界元的功能元素 表达到神经元，并决定它们被移除的发育时间。我们将决定如何 产生SNRPN编码和非编码RNA，并操纵UBE3A-ATS和UBE3A-AT的表达水平 UBE3A以确定它们对UBE3A印记的影响。最后，我们将检验UBE3A-ATS的假设 通过转录干扰机制沉默父亲的UBE3A。透彻了解 UBE3A印迹表达的机制可能揭示新的基因调控机制 适用于基因组的其他地方，并可能确定治疗AS的新的治疗靶点。

SUMMARY Imprinted expression of UBE3A is neuron-specific and occurs because the paternally-inherited allele is silenced. A long non-coding antisense transcript, UBE3A-ATS, is expressed in a neuron-specific manner and mediates UBE3A imprinting by an unknown mechanism. The overall goal of this proposal is to understand the processes underlying the regulation of neuron-specific expression of UBE3A-ATS and repression of paternal UBE3A in human neurons. We propose to investigate three different aspects regulating UBE3A imprinted expression: 1) the regulation of coding versus non-coding SNRPN RNAs in human neurons; 2) the tissue-specific regulation of UBE3A-ATS, the distal portion of the SNRPN ncRNA; and 3) the mechanism by which UBE3A-ATS leads to the repression of paternal UBE3A. We will use patient-specific induced pluripotent stem cells (iPSCs) derived from Angelman syndrome (AS) patients and their neuronal derivatives for these studies. We will dissect the functional elements of the cis-acting boundary element restricting UBE3A-ATS expression to neurons and determine the developmental timing of their removal. We will determine how SNRPN coding versus non-coding RNA is produced and manipulate expression levels of UBE3A-ATS and UBE3A to determine their effect on UBE3A imprinting. Finally, we will test the hypothesis that UBE3A-ATS silences paternal UBE3A via a transcriptional interference mechanism. A thorough understanding of the mechanisms underlying UBE3A imprinted expression may reveal novel gene regulatory mechanisms applicable elsewhere in the genome, and may identify novel therapeutic targets for treating AS.