Regulation of UBE3A Imprinted Expression

UBE3A 印迹表达的调控

基本信息

批准号：
10255508
负责人：
Brenton R. Graveley
金额：
$ 41.66万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10255508
关键词：
Alleles Angelman Syndrome Antisense Oligonucleotides Boundary Elements CRISPR interference Cell Line Cerebrum Chromosomes Clustered Regularly Interspaced Short Palindromic Repeats Code Development Disease Distal Elements Engineering Excision Exons F2R gene Genes Genetic Transcription Genome Genome engineering Goals Human In Vitro Individual Induced pluripotent stem cell derived neurons Inherited Investigation Maps Mediating Neurons Organoids Patients Process Proteins Public Health RNA RNA Polymerase II Regulation Regulator Genes Regulatory Element Repression SNRPN Testing Therapeutic Tissues Transcript Transcriptional Regulation UBE3A gene Untranslated RNA cis acting element engineered stem cells epigenetic silencing experimental study imprint induced pluripotent stem cell monolayer neurogenesis neurogenetics new therapeutic target novel promoter

项目摘要

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.

概括 ube3a的印迹表达是神经元特异性的，并且由于父亲含有的等位基因是沉默。长的非编码反义转录本ube3a-ats以神经元特异性的方式表达通过未知机制介导UBE3A烙印。该提议的总体目标是了解 UBE3A-ATS神经元特异性表达调节的过程和抑制人类神经元中的父亲ube3a。我们建议调查调节UBE3A的三个不同方面印迹表达：1）人神经元中编码与非编码SNRPN RNA的调节； 2） ube3a-ats的组织特异性调节，snrpn ncRNA的远端部分； 3）机制 ube3a-ats导致了对父亲Ube3a的镇压。我们将使用特定于患者的诱导多能源自Angelman综合征（AS）患者及其神经元衍生物的干细胞（IPSC）研究。我们将剖析限制ube3a-ats的顺式作用边界元素的功能元素表达对神经元的表达并确定其去除的发育时机。我们将确定如何生产SNRPN编码与非编码RNA，并操纵UBE3A-ATS的表达水平和 UBE3A确定它们对UBE3A印迹的影响。最后，我们将检验ube3a-ats的假设通过转录干扰机制沉默父亲UBE3A。对 UBE3A表达表达的机制可能揭示新的基因调节机制适用于基因组的其他地方，并可以识别出新的治疗靶标的。