Regulation of UBE3A Imprinted Expression

总结 UBE 3A的印记表达是神经元特异性的，并且发生是因为父系遗传的等位基因是 沉默一种长的非编码反义转录物UBE 3A-ATS以神经元特异性方式表达， 通过未知的机制介导UBE 3A印迹。本提案的总体目标是了解 UBE 3A-ATS的神经元特异性表达的调节和抑制的潜在过程 父亲的UBE 3A在人类神经元中的表达。我们建议研究调节UBE 3A的三个不同方面 印迹表达：1）人类神经元中编码与非编码SNRPN RNA的调节; 2） SNRPN ncRNA的远端部分UBE 3A-ATS的组织特异性调节;以及3）通过 其中UBE 3A-ATS导致父源UBE 3A的抑制。我们将使用患者特异性诱导多能 来源于Angelman综合征（AS）患者的干细胞（iPSC）及其神经元衍生物， 问题研究我们将剖析限制UBE 3A-ATS的顺式作用边界元件的功能元件 表达到神经元，并确定其去除的发育时间。我们将决定如何 产生SNRPN编码RNA与非编码RNA，并操纵UBE 3A-ATS的表达水平， UBE 3A，以确定它们对UBE 3A印迹的影响。最后，我们将检验UBE 3A-ATS 通过转录干扰机制使父本UBE 3A沉默。的透彻理解 UBE 3A印迹表达的潜在机制可能揭示新的基因调控机制 适用于基因组中的其他地方，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.