Gene Regulatory Non-Coding RNAs in the Human Heart

这项由心脏分子遗传学家和 RNA-染色质生物化学家将研究染色质相关， 转录因子依赖的非编码RNA（ncRNA）作为必需的 心脏调节网络的组成部分。我们采用了一种新的方法 用于鉴定功能性长链非编码RNA（lncRNA）， 包括基因调控网络的增强子。利用转录因子- 依赖性，我们确定ncRNA作为顺式调控元件的标记物 对小鼠心律基因调控网络至关重要。这种方法 鉴定出异常强的增强子，它们相关的ncRNA是 染色质结合的和增强子功能所需的。这个主集的证明 研究lncRNA及其相关增强子的阶段 人类心脏基因调控网络。我们支持我们的应用程序 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和顺式调节元件 （克雷斯）对于人心律控制基因的表达是必需的。在 第一个具体目标，我们将询问一组由其TF定义的lncRNA， 依赖性和染色质定位的CRE活性的要求， 靶基因表达。在第二个目标中，我们将利用新的分子工具 为了探索lncRNA调节基因的分子机制， 表情这些相互关联的目标共同依赖于互补和 合作调查人员的专业知识不重叠， ncRNA生物学中的重要问题。这些假设适用于 人类遗传学、转录调控和RNA生物学，因此可以 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.