A genomic approach to studying the life cycle of intron lariats

研究内含子套索生命周期的基因组方法

• 批准号: 10335280
• 负责人: William G Fairbrother
• 金额: $ 43.96万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-16 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10335280
• 关键词: Algorithmic Software; Area Analyses; Biochemical; Biological; Catalysis; Cell Line; Cells; Chromatin; Clinical Medicine; DNA Polymerase II; Data; Defect; Disease; Encephalopathies; Enzymes; Event; Exercise; Exons; Family; Funding; Genes; Genetic; Genetic Transcription; Genome; Genomic approach; Grant; Hereditary Disease; Heritability; Herpesviridae; Herpesvirus 1; Human; Human Genome; Infection; Innate Immune System; Introns; Lead; Libraries; Life; Life Cycle Stages; Link; Maps; Measures; Messenger RNA; Methods; Mining; Mutation; Natural Immunity; Nucleotides; Oncogenic; Paper; Process; Publishing; RNA; RNA Splicing; RNA-Binding Proteins; Recovery; Recycling; Research; Risk; Role; Sampling; Severities; Site; Small Nuclear Ribonucleoproteins; Spliceosomes; System; Techniques; Testing; Transcript; Untranslated RNA; Viral; Work; causal variant; clinical sequencing; deep sequencing; expectation; high reward; in vivo; insight; latency associated transcript; loss of function mutation; mRNA Precursor; member; novel; research study; scale up; signal processing; success

Project Summary/Abstract Pre-mRNA splicing is a critical and regulated processing event where introns are precisely excised from nascent RNA transcripts. As many as one third of all heritable disease mutations result in splicing defects. This research studies the role of branchpoints in determining splice site selection (3'ss) is utilized in vivo and also the effect branchpoints have on the life cycle of the intron. Each pre-mRNA splicing event creates a lariat and spliced exon junction. While a great deal is known about splice exon junctions almost nothing is known about lariats. By mapping all branchpoints in the human genome, we are opening up a whole new area of analysis. The identification of branchpoints by transcript data will facilitate the interpretation of clinical sequencing data. In addition to the intrinsic value of this data, the successful completion of this proposal will test some hypothesis about the fundamental catalysis and recognition that occurs in vivo in the processing of eukaryotic genes. Studying these intermediates at a system wide level will bring a biochemical-level understanding to hundreds of thousands of processing events. Furthermore, each intron lariat has a lifecycle – created by splicing of a transcribed product, recycled by debranching and degradation. The recycling of introns is vital to replenishing the intracellular levels of free nucleotides and to return splicing factors into active spliceosomes. Some introns have a second life after splicing as non- coding RNAs (ncRNAs). As we are sampling steady state levels of introns we gain insight into both these processes. This proposal seeks to follow this lead by exploring some of the reasons why certain introns appear stabilized.

项目总结/摘要 前体mRNA剪接是一个关键的和受调控的加工事件，其中内含子被精确地 从新生RNA转录物中切除。在所有遗传性疾病的突变中， 导致拼接缺陷。本研究探讨了分支点在决定剪接中的作用 位点选择（3 'ss）在体内被利用，并且分支点对生命周期的影响也被利用。 内含子每个前体mRNA剪接事件产生一个剪接和剪接的外显子连接。而 关于剪接外显子连接知道很多，关于剪接外显子连接几乎一无所知。通过 绘制人类基因组的所有分支点，我们正在开辟一个全新的领域， 分析.通过转录本数据识别分支点将有助于解释 临床测序数据。除了这些数据的内在价值， 这一提议的一部分将检验一些关于基本催化和认识的假设， 发生在体内真核基因的加工过程中。在一个系统中研究这些中间体 广泛的水平将带来生物化学水平的理解，以数十万的处理 事件此外，每个内含子lxl都有一个生命周期-通过剪接转录的 产物，通过脱支和降解再循环。内含子的再循环对于 补充游离核苷酸的细胞内水平，并使剪接因子恢复活性， 剪接体一些内含子在剪接成非编码RNA（ncRNA）后有第二次生命。 当我们对内含子的稳态水平进行采样时，我们可以深入了解这两个过程。 这项提案试图通过探索某些内含子在基因组中的作用的原因来跟踪这一线索。 看起来很稳定。