GENOMIC RESPONSES TO HORMONE SIGNALING

在任何给定的细胞中，只有所有编码基因的子集表达，并且 表达的水平和时机支配着细胞的发育， 分化、功能和生理。在这里，我们描述了一种用于 多个基因表达水平的同时监测 平行的。该方法涉及到确定相对的 基于整个mRNA杂交的mRNAs浓度 人工合成寡核苷酸高密度阵列的种群 (GeneChips TM)。这里提出的实验所使用的阵列 包含超过65,000种不同的25聚体寡核苷酸 在第一阶段，我们将：1)设计一套 用于同时定量超过6500个的阵列(GeneChip TM) 人类基因(Hum6000)，来自GenBank和公共EST数据库(DBEST)。 2)证明该方法具有灵敏、特异、定量的特点。 3)使用Hum6000芯片确定动态调节的mRNA 三种激素受体(β2肾上腺素能、CCR5、催产素)代表 三条不同的信号通路。在第二阶段，我们将使用Hum6000 芯片用于分析受大量基因动态调节的mRNA 受体面板和药理探针。我们还将确认一项 通过使用Northern分析来自Hum6000芯片的发现的子集， 蛋白质印迹和基因表达标记物。 建议的商业应用： 对大量基因表达水平的定量监测 基因应该被证明在阐明基因功能、探索 疾病的原因和机制，以及发现潜在的 治疗和诊断目标。作为基因组信息的主体 增长，此处描述的类型的高度并行的方法提供 高效、直接地利用序列信息来阐明 重要的激素、药物和代谢途径。新技术的发展 基因芯片技术作为监测全基因组表达谱的工具 具有多种生物医学、制药和农业应用。 收入将来自出售基因芯片系统(流体) 站和扫描仪)和定制激素受体的GeneChips研究 将确定可能成为药物新靶点的基因 发展。荷尔蒙激活基因的调节区可能 还可用于开发快速、非放射性的高放射性报告分析方法。 对激活激素受体的药物进行全量筛选。

Only a subset of all encoded genes is expressed in any given cell, and the levels and timing of expression govern cellular development, differentiation, function and physiology. Here we describe a method for the simultaneous monitoring of the expression levels of many genes in parallel. The method involves the determination of the relative concentrations of mRNAs based on hybridization of entire mRNA populations to high-density arrays of synthetic oligonucleotides (GeneChips TM). The arrays used for the experiments proposed here contain more than 65,000 different 25-mer oligonucleotides of defined sequence in an area of 1.6 cm 2. In Phase I, we will: 1) Design a set of arrays (GeneChip TM) for the simultaneous quantitation of over 6500 human genes (Hum6000) from GenBank and the public EST database (dbEST). 2) Demonstrate that the method is sensitive, specific and quantitative. 3) Use the Hum6000 chip to determine the mRNAs dynamically regulated by three hormone receptors (beta-2 Adrenergic, CCR5, Oxytocin) representing three different signaling pathways. In Phase II we will use the Hum6000 chip to analyze the mRNAs that are dynamically regulated by a large panel of receptors, and pharmacologic probes. We will also confirm a subset of the findings from the Hum6000 chip by using Northern analysis, Western blots, and gene expression markers. PROPOSED COMMERCIAL APPLICATION: The quantitative monitoring of expression levels for large numbers of genes should prove valuable in elucidating gene function, exploring the causes and mechanisms of disease, and for the discovery of potential therapeutic and diagnostic targets. As the body of genomic information grows, highly parallel methods of the type described here provide an efficient and direct way to use sequence information to elucidate important hormone, drug and metabolic pathways. Development of the GeneChip technology as a tool to monitor genome-wide expression profiles has multifold biomedical, pharmaceutical and agricultural applications. Revenue will be derived from the sale of the GeneChip system (fluidics station and scanner) and custom GeneChips Studies with hormone receptors will identify genes that will be potential new targets for drug development. The regulatory regions of hormonally activated genes could also be used to develop rapid, non-radioactive reporter assays for high- throughput screening of drugs that activate hormone receptors.