A genomic approach to studying the life cycle of intron lariats

前-mRNA剪接是一个关键的和受调控的加工事件，其中内含子 精确地从新生的RNA转录本中切下。多达三分之一的可遗传疾病 突变会导致剪接缺陷。本研究研究了分支点在生物信息学中的作用 确定剪接位点选择(3‘s)在体内的应用以及分支点的影响 关于内含子的生命周期。每个前mRNA剪接事件都会产生套索和剪接外显子 交叉口。虽然人们对剪接外显子连接的了解很多，但几乎一无所知 关于拉尼尔斯。通过绘制人类基因组中的所有分支点，我们正在打开一个 全新的分析领域。通过文字记录数据识别分支点将 促进对临床测序数据的解释。除了它的内在价值 这些数据，这一提议的成功完成将检验关于 在真核基因处理过程中在体内发生的基本催化和识别。 在系统范围的水平上研究这些中间体将带来对生物化学水平的理解 数十万个处理事件。此外，每个内含子套索都有一个生命周期- 通过剪接转录产物产生的，通过去分支和降解来回收的。这个 内含子的循环对于补充细胞内游离核苷酸水平和 将剪接因子返回到活跃的剪接体。一些内含子在剪接后有第二次生命 作为非编码RNA(NcRNAs)。当我们采样内含子的稳态水平时，我们获得了洞察力 在这两个过程中。这项提议试图通过探索一些原因来追随这一线索 为什么某些内含子看起来很稳定。

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.