Practical High Througput Low Cost Gene Expression Analysis

项目概述/摘要：本项目拟开发和应用一种基于高通量、多路聚合酶链反应（pcr）的基因表达分析方法，使基因表达谱分析广泛应用于药物研究、开发和高含量筛选。拟议的项目建立在使用两项关键技术的基础上，Althea的多路XP PCR和blushift的IsoCyte高性能平板扫描仪。这两种技术结合在可行性研究中，通过使用一种新方法来制备用于监测基因表达端点的条形码随机头阵列。最终目标是一种在2分钟内扫描1536个样本，对每个样本进行20-40个基因的多路基因表达分析的方法。项目简介：破译癌症、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。这种疾病遗传学知识库的指数级增长导致了大量基因、蛋白质和途径的识别，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目标。现在的挑战是通过实验更深入地研究这些基因在疾病中如何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.