Standardized Gene Expression Core Facility Development

描述(由申请人提供)： 需要灵敏、高通量、标准化的基因表达 俄亥俄州医学院针对癌症的特异性分析 总体来说，研究社区。基因表达之间的经验关联性 模式和临床上重要的表型将提供改进的标记 预测癌症行为和对特定药物治疗的反应。在……里面 此外，这种协会将为新的机制提供一个知情的基础 旨在改善癌症预防、诊断和治疗的研究。 微阵列(cDNA和/或寡核苷酸)基因表达分析是一种 有价值的筛查工具，为此，俄亥俄州医学院 目前正在建立微阵列核心设施。 然而，很明显，微阵列分析不够灵敏， 可重复性或灵活性，可用于许多临床应用。因此， 微阵列数据一般必须在后续的定量基因中进行验证 表情分析。由六个NCI组成的小组赞助了 俄亥俄医学院加入支持这一申请，以建立一个 高通量、标准化的基因表达分析设备 标准化RT(START)-PCR。这将是一种全新的资源，这 小组将与其他机构的调查人员一起参加AIM 4，他们是 使用微阵列，并支持使用启动-聚合酶链式反应作为“黄金标准” 他们可以优化微阵列分析协议。这个设施将是 以较低的成本向一般研究团体提供。高潮 启动型聚合酶链式反应的重复性、灵敏度和低检测阈值 基因表达方法已经在多个独立研究中得到验证。在……里面 在一项盲法研究中，在一个普通的cdna中测量了10个基因的表达。 实验室。实验室间的平均方差相关系数(CV)为 0.48，证实基因表达可能存在2-3倍的差异 信心十足地被检测到。利用微通道毛细管电泳法 降至0.26。这项建议的具体目标如下。I； 建立4,000个基因的自动化START-PCR核心设施 表达检测/24小时。目标2；以低成本向 NCI资助的MCO调查人员以及其他感兴趣的调查人员 机构，3；开发一个带注释的基因表达数据库，为两者提供服务 NCI资助的MCO调查人员以及其他机构，AIM：4开发 一种可接受的标准化微阵列数据分析的方法。

DESCRIPTION (provided by applicant): There is a need for sensitive, high throughput, standardized gene expression analysis at the Medical College of Ohio specifically and for the cancer research community in general. Empiric association between gene expression patterns and clinically important phenotypes will provide improved markers to predict cancer behavior and response to treatment with specific agents. In addition, such associations will provide an informed basis for new mechanistic studies aimed at improving cancer prevention, diagnosis and treatment. Microarray (cDNA and/or oligonucleotide) gene expression analysis is a valuable screening tool and for this reason, the Medical College of Ohio currently is in the process of establishing a microarray core facility. However, it is clear that microarray analysis is not sufficiently sensitive, reproducible, or flexible for many clinical applications. Consequently, microarray data generally must be validated in follow-up quantitative gene expression analyses. The group of six NCI sponsored investigators at the Medical College of Ohio join in supporting this application to establish a high throughput, standardized gene expression analysis facility based on standardized RT (StaRT)-PCR. This will be a completely new resource, This group will be joined in Aim 4 by investigators at other institutions who are using microarrays and support the use of StaRT-PCR as a "gold standard" by which they may optimize microarray analysis protocols. This facility will be available to the general research community at low cost. The high reproducibility, sensitivity, and low detection threshold of the StaRT-PCR gene expression method have been validated in multiple independent studies. In a blinded study, expression of ten genes was measured in a common cDNA by four laboratories. The average inter-laboratory correlation of variance (CV) was 0.48, confirming that 2-3 fold differences in gene expression could be detected with confidence. Using microchannel capillary electrophoresis the CV was reduced to 0.26. The Specific Aims of this proposal are as follows. I; establish an automated StaRT-PCR core facility capable of 4,000 gene expression assays/24 hours. AIM 2; Provide this service at low cost to the NCI-funded investigators at MCO as well as interested investigators at other institutions, 3; Develop an annotated gene expression database to service both NCI-funded investigators at MCO as well as other institutions, ,AIM: 4 develop an acceptable method for standardizing microarray data analysis.