Large-scale characterization of the function of RNA regulatory elements

RNA调控元件功能的大规模表征

• 批准号: 10487581
• 负责人: Eric Lyman Van Nostrand
• 金额: $ 48万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10487581
• 关键词: Address; Affect; Antisense Oligonucleotides; Area; Binding Sites; Biological Assay; Biology; Catalogs; Cell Nucleus; Databases; Development; Disease; Elements; Foundations; Genetic Variation; Genomic approach; Genomics; Half-Life; Human; Human Genetics; Individual; Knowledge; Location; Maps; Mediation; Methods; Pharmaceutical Preparations; Physiological; Proteins; RNA; RNA Binding; RNA Decay; RNA Processing; RNA Splicing; RNA Stability; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Regulatory Element; Reporter; Research; Research Personnel; Ribosomes; Site; Small Nuclear RNA; Small Nucleolar RNA; Spinal Muscular Atrophy; Techniques; Transcript; Translations; Work; improved; insight; interest; knock-down; tool

PROJECT SUMMARY Once an RNA is transcribed in the nucleus, it is bound by RNA binding proteins and other regulatory RNAs which control its sequence (by altering splicing), half-life (by modulating RNA stability and decay) and translation (through mediation of ribosome initiation and elongation) among other RNA processing steps. Individual RBP binding sites can have significant physiological importance, as emphasized by the recent example of the drug Spinraza, which is an antisense oligonucleotide that blocks a single RBP:RNA interaction in order to cure Spinal Muscular Atrophy. Recent advances in genomics techniques have dramatically increased our ability to identify the interaction sites for these regulatory RBPs and RNAs, and we now have catalogs of over a million such interaction sites that are candidate RNA regulatory elements. However, it is clear that only a small fraction of these elements truly function as regulatory modules, as few show differences in RNA processing when the RBP is knocked down or otherwise altered. As such, large-scale assays to sift through these elements to identify the subset that are function are an essential missing piece in converting these element databases into a useful tool for researchers interested in understanding whether human genetic variation will alter RNA biology. In this proposal, I describe my research group’s proposed efforts to address this major knowledge gap in two areas: 1. Using orthogonal approaches (rapid degradation of RBPs followed by genomics profiling to identify direct regulatory targets of RBPs and massively parallel reporter assays) to characterize which RBP binding sites confer regulation. 2. Large-scale identification of regulatory targets for snoRNAs, snRNAs, and other regulatory RNAs through improved genomics techniques. My extensive expertise in developing experimental and computational genomics methods to map and understand RNA processing regulatory networks makes my newly founded lab an ideal location to undertake these efforts, and build an improved global picture of the RNA processing regulatory landscape. Further, it will support my efforts to develop an independent research group that will lay the foundation to the broader effort to understand how human genetic variation affects disease through mis-regulation of RNA processing.

项目摘要 一旦将RNA转录在细胞核中，它就会由RNA结合蛋白和 其他调节性RNA控制其序列（通过更改剪接），半衰期（通过调节 RNA稳定性和衰减）和翻译（通过核糖体计划的调解和 伸长）除其他RNA处理步骤外。单独的RBP绑定站点可以具有 正如最近的药物例子所强调的那样，身体上的重要性很大 Spinraza，这是一种反义寡核苷酸，可阻止单个RBP：RNA相互作用 为了治愈脊柱肌肉萎缩。基因组技术的最新进展具有 大大提高了我们确定这些调节RBP的相互作用位点的能力和 RNA，我们现在拥有超过一百万此类互动站点的目录，这些网站是候选RNA 监管要素。但是，很明显，这些元素中只有一小部分 充当调节模块，因为RBP为 撞倒或以其他方式改变。因此，通过这些元素进行大规模测定 识别功能的子集是转换这些的必不可少的一部分 对于有兴趣了解人类的研究人员，元素数据库成为有用的工具 遗传变异将改变RNA生物学。 在此提案中，我描述了我的研究小组提出的解决这一专业的努力 在两个领域的知识差距： 1。使用正交方法（RBP的快速降解，然后是基因组学 分析以识别RBP和大规模平行报告的直接调节目标 分析）以表征哪些RBP绑定站点会议法规。 2。大规模识别snornas，snrnas和其他的调节目标 通过改进基因组技术的调节RNA。 我在开发实验和计算基因组学方法方面的广泛专业知识 地图并了解RNA处理调节网络使我的新成立的实验室成为 进行这些努力并建立RNA的全球图景的理想位置 处理监管景观。此外，它将支持我建立独立的努力 研究小组将为了解人类的更广泛努力奠定基础 遗传变异通过RNA处理的错误调节影响疾病。