Practical High Througput Low Cost Gene Expression Analysis

描述（由申请人提供）：项目摘要/摘要：这里提出的是一个高通量，多重PCR为基础的方法，基因表达分析的开发和应用，以广泛地使基因表达谱用于药物研究，开发和高内容筛选。拟议项目建立在使用2项关键技术的基础上，Althea的多重XP PCR和Blueshift的IsoCyte高性能平板扫描仪。在拟定的可行性研究中，通过使用一种新方法制备用于监测基因表达终点的条形码随机微珠阵列，将这2种技术结合在一起。最终目标是一种用于每个样品20-40个基因的多重基因表达分析的方法，其扫描通量为2分钟内1536个样品。项目叙述：癌症、糖尿病、心血管疾病和炎症性疾病等复杂疾病的遗传学被破译的速度一直在加快。研究人员已经开始详细描述导致这些复杂疾病的多种遗传机制，以及与疾病反应相关的许多功能途径，如应激和损伤反应，免疫反应，细胞周期调节，细胞增殖和细胞死亡。我们的疾病遗传学知识库的这种指数增长导致了大量基因、蛋白质和途径的鉴定，这些基因、蛋白质和途径可能在疾病发展中发挥核心作用，并可能成为治疗干预的潜在靶点。现在的挑战是通过实验更深入地研究这些基因如何在疾病中发挥病理和保护作用，以及不同的化合物和化合物类别如何影响这些基因。

DESCRIPTION (provided by applicant): Project Summary/Abstract: Proposed here is the development and application of a high-throughput, multiplexed PCR-based approach to gene expression analysis to broadly enable the use of gene expression profiling for drug research, development and high content screening. The proposed project builds off of the use of 2 key technologies, Althea's multiplexed XP PCR, and Blueshift's IsoCyte high performance plate scanner. These 2 technologies are combined in the proposed feasibility study through the use of a novel approach to preparing a barcoded random bead array for monitoring the gene expression endpoints. The end goal is a method for the performance of multiplexed gene expression analysis of 20-40 genes per sample with a scanning throughput of 1536 samples in 2 minutes. Project Narrative: The pace at which the genetics of complex diseases, such as cancer, diabetes, cardiovascular and inflammatory disorders, are being deciphered has been accelerating. Researchers have begun to characterize in detail multiple genetic mechanisms that give rise to these complex diseases, as well as numerous functional pathways associated with disease response such as stress and damage response, immune response, cell cycle regulation, cell proliferation and cell death. This exponential growth in our knowledgebase of disease genetics has led to the identification of a large array of genes, proteins and pathways that potentially play a central role in disease development and may be potential targets for therapeutic intervention. The challenge now is to experimentally delve deeper, both into how these genes may function pathologically and protectively in disease, and into how different compounds and compound classes might influence these genes.