High-Throughput DNA Microarrays for Inflammation

描述（申请人提供）：一种定量高通量基因芯片 基于核酸酶保护的ArrayPlateTM检测方法已被开发用于药物 的发现我们将利用这项技术开发两种新的检测方法， 筛选影响转录因子表达的化合物， 参与炎症的调节基因。任何表达的微小变化 将对暴露于推定的候选药物的细胞的基因进行定量。 一种测定将定量两种参与转录的特异性转录因子， 炎症和它们调节的几个基因（共16个）。另一个将测试 一组广泛的与免疫相关的转录因子的表达 与控制其他生理途径的因素一起响应（300 共计）。将筛选2000种新型甾体化合物的模型库。 ArrayPlateTM允许DNA阵列技术用于高通量- 准确度和精确度。细胞在核酸酶保护下裂解 寡核苷酸，其杂交并保护所有靶向mRNA。后 核酸酶消化寡核苷酸化学计量地结合到靶向 在基因芯片上对序列进行定量。杂交特异性得到确认 在每一个样品中。该方法足够灵敏，可以测量非常罕见的mRNA- 从96孔板中生长的30，000个细胞中，每个细胞一个mRNA分子。

DESCRIPTION (provided by applicant): A quantitative high-throughput gene chip assay, ArrayPlateTM, based on nuclease protection has been developed for drug discovery. We will use this technology to develop two novel assays designed to screen for compounds that affect the expression of transcription factors and regulated genes involved in inflammation. Small changes in expression for any of the genes will be quantified for cells exposed to putative drug candidates. One assay will quantify two specific transcription factors involved in inflammation and several genes they regulate (16 total). Another will test expression of a wide panel of transcription factors implicated in the immune response alongside factors controlling other physiological pathways (300 total). A model library of 2000 novel steroidal compounds will be screened. ArrayPlateTM allows DNA array technology to be used in high throughput - with accuracy and precision. Cells are lysed in the presence of nuclease protection oligonucleotides, which hybridize to and protect all targeted mRNAs. After nuclease digestion oligonucleotides bound stochiometrically to targeted sequences are quantified on gene chips. Hybridization specificity is confirmed within each sample. The method is sensitive enough to measure very rare mRNAs - down to one mRNA molecule per cell, from 30,000 cells grown in a 96-well plate.