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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.

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 通过转录干扰机制沉默父本UBE 3A。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。