Gene Regulatory Non-Coding RNAs in the Human Heart

这项由心脏分子遗传学家和一个 RNA-染色质生物化学家将研究与染色质相关的 转录因子依赖的非编码RNA(NcRNAs)是必需的 心脏调节网络的组成部分。我们应用了一种新的方法 用于鉴定功能较长的非编码RNA(LncRNAs)和 构成基因调控网络的增强剂。使用转录因子- 依赖，我们发现ncRNAs是顺式调节元件的标志 对小鼠心律基因调控网络至关重要。这种方法 确定了特别强的增强子，及其相关的ncRNA是 染色质结合，是增强功能所必需的。主集的这一证明 LncRNAs及其相关增强子的研究阶段 人类心脏基因调控网络。我们假设我们的应用程序 人类心肌细胞的方法将允许识别一个功能类别 人类心脏增强子相关的lncRNA和顺式调节元件 (Cres)对人类心律控制基因的表达至关重要。在……里面 第一个具体目标，我们将询问一组由其Tf-Tf定义的LncRNA- 对Cre活性和Cre活性需求的依赖性和染色质定位 靶基因表达。在第二个目标中，我们将利用新的分子工具 探讨lncRNAs调控基因的分子机制 表情。这些相互关联的目标共同依赖于互补和 合作调查人员的非重叠专业知识解决了高度 NcRNA生物学中的重大问题。这些假设适用于 人类遗传学、转录调控和RNA生物学，因此可能 在心血管遗传学和更广泛的人类中都有影响 分子遗传学。

This collaborative proposal from a cardiac molecular geneticist and a RNA-chromatin biochemist will investigate chromatin associated and transcription factor dependent non-coding RNAs (ncRNAs) as essential components of a cardiac regulatory network. We have applied a novel approach for identification of functional long non-coding RNAs (lncRNAs) and the enhancers that comprise gene regulatory networks. Using transcription factor- dependence, we identified ncRNAs as markers of cis-regulatory elements essential for a mouse cardiac rhythm gene regulatory network. This approach identified exceptionally strong enhancers, and their associated ncRNA are chromatin-bound and required for enhancer function. This proof of principal sets the stage for the investigation of lncRNAs and their associated enhancers in human cardiac gene regulatory networks. We posit that application of our approach to human cardiomyocytes will allow identification of a functional class of human heart enhancer-associated lncRNAs and cis-regulatory elements (CREs) essential for the expression of human cardiac rhythm control genes. In the first specific aim, we will interrogate a set of lncRNAs defined by their TF- dependence and chromatin localization for their requirement for CRE activity and target gene expression. In the second aim, we will utilize novel molecular tools to probe the molecular mechanisms whereby the lncRNAs modulate gene expression. Together these interconnected aims rely on the complementary and non-overlapping expertise of the collaborative investigators to address a highly significant problem in ncRNA biology. These hypotheses are applicable to human genetics, transcriptional regulation, and RNA biology, and therefore may have impact both within cardiovascular genetics and more broadly within human molecular genetics.