长链非编码RNA与蛋白相互作用在胚胎干细胞多能性中的研究

Human and mouse genomes encode large numbers of long noncoding RNAs (lncRNAs), which are involved in diverse biological processes, such as imprinting. However, molecular and biological functions of lncRNAs are largely unknown. As no naked RNAs are present inside cells，RNA must function through interaction with proteins. Thus, understanding of RNA-protein interaction in vitro may infer lncRNA function in vivo. In this study, we propose to profile RNA-protein interactions in a global and high-throughput manner. We have selected a number of lncRNAs highly or specifically expressed in embryonic stem cells (ESCs). To uncover their function, we will first probe each lncRNA transcript on a HuProt array, which contains over 17,000 unique proteins, to comprehensively and unbiasedly reveal all proteins interacting with a lncRNA transcript. LncRNA-protein interactions will be validated in vitro and in vivo and further investigated for their biological importance in the context of ESC pluripotency. Construction of a two-dimensional RNA-protein regulatory network will facilitate our understanding of lncRNAs in regulating cellular functions, including signal transduction and epigenetic regulation of gene expression, and provide novel insights in how lncRNAs function in ESC self-renewal and differentiation.

人类和鼠的基因组编码大量的长链非编码RNAs（lncRNAs），它们在遗传印记等多种生物学过程中起到重要作用。然而，大多数lncRNAs的生物学功能和分子机制尚不清楚。本项目通过高通量筛选与胚胎干细胞（embryonic stem cells,ESCs）中lncRNAs相互结合的蛋白，来阐明lncRNAs在干细胞多能性中的分子生物学功能。首先，我们将利用包含1.7万个人类蛋白的全蛋白组芯片，对ESCs特异表达的lncRNAs-蛋白相互作用进行大规模的、全局性的分析和筛选。并在体内、外利用蛋白和RNA双向互作实验来验证以上蛋白组规模的相互作用，构建RNA-蛋白二维调控网络。最后，研究lncRNAs及它与蛋白互作的生物学功能。研究成果将阐明lncRNAs发挥生物学功能的分子机制，促进我们对lncRNA在表观遗传调控中作用的认识，为研究lncRNA在胚胎干细胞自我更新和分化中的作用提供新思路。