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

基本信息

批准号：
10155500
负责人：
William G Fairbrother
金额：
$ 59.24万
依托单位：
BROWN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-16 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10155500
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 Structure 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

项目摘要

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.

项目概要/摘要前 mRNA 剪接是一个关键且受调控的加工事件，其中内含子精确地多达三分之一的遗传性疾病突变被从新生 RNA 转录本中切除。导致拼接缺陷。这项研究研究了分支点在确定拼接中的作用。体内利用位点选择（3'ss），并且分支点对生命周期的影响每个前mRNA剪接事件都会产生一个套索和剪接的外显子连接。人们对剪接外显子连接了解甚多，但对套索却一无所知。绘制人类基因组中的所有分支点，我们正在开辟一个全新的领域通过转录数据识别分支点将有助于解释。除了这些数据的内在价值之外，成功完成的临床测序数据。该提案的一部分将检验一些关于基本催化和认识的假设在系统中研究这些中间体发生在体内真核基因的加工过程中。广泛的水平将为数十万个处理带来生化水平的理解此外，每个内含子套索都有一个生命周期——通过转录的剪接而创建。内含子的回收对于脱支和降解至关重要。补充细胞内游离核苷酸的水平并使剪接因子恢复活性一些内含子在剪接后作为非编码 RNA (ncRNA) 具有第二次生命。当我们对内含子的稳态水平进行采样时，我们深入了解了这两个过程。该提案旨在通过探索某些内含子的一些原因来遵循这一先导显得稳定。