Genomic Measurement of Alternative Splicing by DNA Array

描述（由申请人提供）：选择性剪接是哺乳动物基因表达控制的关键过程，也是蛋白质多样性的主要来源。 剪接调控中的错误涉及许多疾病过程，包括癌症，但参与这种调控的细胞回路大多是未知的。利用DNA微阵列进行基因表达谱分析，使我们在全基因组范围内对基因调控系统的理解取得了重大进展。然而，没有一个常用的微阵列分析系统可以检测到剪接的变化，而是提供每个基因的整体转录丰度的信息。这些方法提供了第一层基因调控的信息，但错过了许多重要的变化，基因产物引起的剪接模式的改变。这里提出的项目将使三个小组合作建立一种新型的DNA微阵列，允许并行分析多种选择性剪接模式。此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。我们将设计和生产寡核苷酸阵列，检测和测量小鼠中的剪接事件。这种微阵列方法是建立良好的酵母剪接，最近的结果表明，它能够检测更复杂的人类细胞系和组织中的选择性剪接。这里的建议是将其应用于小鼠的特定调节系统，包括神经系统和心脏。该项目的目标是将剪接调控的研究推向全基因组水平，全面寻找受特定剪接调控途径影响的基因，并为基因表达谱提供更高的精度。

DESCRIPTION (provided by applicant): 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.