Gene Regulatory Non-Coding RNAs in the Human Heart

这项合作提案由一位心脏分子遗传学家和一位 RNA-染色质生物化学家将研究染色质相关和 转录因子依赖性非编码 RNA (ncRNA) 至关重要 心脏调节网络的组成部分。我们采用了一种新颖的方法 用于鉴定功能性长非编码 RNA (lncRNA) 和 组成基因调控网络的增强子。使用转录因子- 依赖性，我们将 ncRNA 鉴定为顺式调控元件的标记 对于小鼠心律基因调控网络至关重要。这种做法 鉴定出异常强的增强子，及其相关的 ncRNA 染色质结合并且是增强子功能所必需的。本金证明集 lncRNA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功能类别 人类心脏增强子相关lncRNA和顺式调控元件的研究 （CRE）对于人类心律控制基因的表达至关重要。在 第一个具体目标，我们将询问一组由其 TF-定义的 lncRNA CRE 活性的依赖性和染色质定位 目标基因表达。在第二个目标中，我们将利用新颖的分子工具 探究lncRNA调节基因的分子机制 表达。这些相互关联的目标共同依赖于互补性和 合作研究人员的专业知识不重叠，以解决高度 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 对心血管遗传学和更广泛的人类都有影响 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.