The role of enhancer RNAs in neuronal plasticity and neurogenesis

增强子RNA在神经元可塑性和神经发生中的作用

• 批准号: 389578377
• 负责人: Professor Dr. Gerrit Begemann
• 金额: --
• 依托单位: Lehrstuhl für Tierphysiologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/389578377?language=en
• 关键词: role; enhancer; RNAs; neuronal; plasticity

Changes in the transcriptional programs of neurons or neuronal progenitors lie at the heart of neuronal plasticity and neurogenesis, respectively. Genes that regulate these processes are under the control of enhancer elements that are differentially regulated during neuronal stimulation and brain development. Enhancers are DNA sequences bound by at times multiple transcription factors. In a combinatorial activation process the combined activity of all bound factors is believed to determine the extent of target gene activation. In addition, neuronal enhancers were recently shown to be actively transcribed, giving rise to non-coding enhancer RNAs (eRNAs). eRNAs significantly contribute to the activation of most immediate early genes in mouse neurons. Moreover, eRNA expression levels serve as a better indicator of enhancer activity than transcription factor binding data. However, how enhancer RNAs are able to direct brain development and neuronal plasticity in vivo is unknown. In particular, it is unknown how much enhancer transcription contributes to enhancer function during these processes. We pursue the hypothesis that enhancer RNAs directly influence the RNA polymerase II (Pol II) transcription machinery by diminishing promoter-proximal pausing of important neuronal genes. To examine this hypothesis, we will combine in vitro biochemistry, next-generation sequencing and in vivo studies of zebrafish neurogenesis. Specifically, we will address the following questions: 1. Are neuronal enhancer RNAs able to abrogate promoter-proximal pausing in vitro? 2. Are neuronal enhancer RNAs conserved between zebrafish and mouse? 3. What is the role of enhancer RNAs in zebrafish nervous system development?Zebrafish is a uniquely suited model system to study eRNA function on an organismic level. It combines the ease of being easily able to image neurogenesis with the ability to manipulate eRNA functionality using both antisense techniques and CRISPRi. Despite the fact that most emerging classes of non-coding RNA influence gene expression on the transcriptional level there is very little insight to date on the underlying mechanisms. This proposal bridges this gap by directly integrating a mechanistic study on Pol II transcription elongation with functional analyses of zebrafish neurogenesis in vivo.

神经元或神经元祖细胞的转录程序的变化分别位于神经元可塑性和神经发生的核心。调节这些过程的基因在神经元刺激和大脑发育过程中受差异调节的增强子元素的控制。增强子是受到多个转录因子结合的DNA序列。在组合激活过程中，所有结合因子的组合活性都被认为确定靶基因激活的程度。此外，最近显示神经元增强剂被积极转录，从而引起非编码增强子RNA（ERNAS）。 ERNAS显着有助于小鼠神经元中最直接的早期基因的激活。此外，与转录因子结合数据相比，ERNA表达水平是增强剂活性的更好指标。但是，增强子RNA如何能够指导脑发育和体内神经元可塑性。特别是，在这些过程中，增强子转录有多少有助于增强子功能。我们提出的假设是，增强子RNA直接通过减少重要神经元基因的启动子 - 抗抑制剂暂停而直接影响RNA聚合酶II（POL II）转录机械。为了审查这一假设，我们将结合体外生物化学，下一代测序和斑马鱼神经发生的体内研究。具体来说，我们将解决以下问题：1。神经元增强子RNA是否能够消除体外启动子暂停的暂停？ 2。在斑马鱼和小鼠之间保守神经元增强子RNA吗？ 3。增强子RNA在斑马鱼神经系统发育中的作用是什么？斑马鱼是一种独特的模型系统，用于研究ERNA在有机水平上的功能。它结合了能够轻松成像神经发生的便利性，并使用反义技术和CRISPRI操纵ERNA功能的能力。尽管大多数新兴的非编码RNA类别在转录水平上影响基因表达，但迄今为止，对基本机制的见解很少。该提议通过直接整合有关POL II转录伸长的机理研究，并在体内对斑马鱼神经发生的功能分析来弥合这一差距。