Genomic Measurement of Alternative Splicing by DNA Array

选择性剪接是控制哺乳动物基因表达的关键过程，也是基因表达的主要来源 蛋白质多样性。剪接调控的错误与包括癌症在内的许多疾病过程有关， 但参与这一调控的细胞电路大多是未知的。利用DNA进行基因表达谱分析 微阵列使我们在理解全基因组上的基因调控系统方面取得了重大进展。 比例。然而，常用的微阵列分析系统都不能检测到剪接的变化 取而代之的是提供每个基因的整体转录丰度的信息。这些方法提供了 关于基因调控的第一层的信息，但错过了由 拼接模式的变化。这里提出的项目将使三个小组能够合作 建立一种可并行分析多种选择性剪接的新型DNA微阵列 模式。这次合作中的每个实验室都在剪接研究方面拥有独特的专业知识。我们将设计 并产生检测和测量小鼠剪接事件的寡核苷酸阵列。这种微阵列 酵母剪接的方法已经建立得很好了，最近的结果表明它有能力检测替代 在更复杂的人类细胞系和组织中进行剪接。这里的建议是将其应用于特定的系统 老鼠的调节，包括神经系统和心脏。该项目的目标是将 研究剪接调控到全基因组水平，全面寻找受 特定的剪接调控途径，并为基因表达谱提供更高的精度。

Alternative splicing is a key process in the control of mammalian gene expression and a major source of protein diversity. Errors in splicing regulation are implicated in many disease processes including cancer, but the cellular circuitry involved in this regulation is mostly unknown. Gene expression profiling using DNA microarrays has led to major advances in our understanding of gene regulatory systems on a genome wide scale. However, none of the commonly available systems of microarray analysis can detect changes in splicing and instead give information on the overall transcript abundance from each gene. These methods provide information on the first layer of gene regulation but miss many important changes in gene product caused by alterations in splicing pattern. The project proposed here will enable three groups to collaborate on establishing a new type of DNA microarray that allows the parallel analysis of multiple alternative splicing patterns. Each of the labs in this collaboration has unique expertise in the study of splicing. We will design and produce arrays of oligonucleotides that detect and measure splicing events in the mouse. This microarray method is well established for yeast splicing, and recent results demonstrate its ability to detect alternative splicing in more complex human cell lines and tissues. The proposal here is to apply it to specific systems of regulation in the mouse, including the nervous system and the heart. The goals of the project are to move the study of splicing regulation to the level of the whole genome, to comprehensively search for genes subject to specific splicing regulatory pathways, and to provide greater precision to gene expression profiling.