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MICROARRAYS TO SCREEN REGULATORY GENES

机译:筛选规则基因的微阵列

摘要

Microarray technology allows the multiple parallel processing of information generated from matrices of huge numbers of loci on a solid substrate, which is useful in the gathering of gene signatures defining specific biological states. An approach has been developed to facilitate this process wherein genes of the same regulatory modality are selected. The transcriptional regulation of these genes is related to the same control element. Primers specific for the regulatory genes are selected, based on minimum cross-reactivity with other genes, using known gene data banks. PCR products of selected regions of known genes either binding to this sequence or whose expression is dependent on this binding, as well as genes interacting with the regulatable genes and control genes, referred to as "amplicons" or "gene cDNA fragments" of between about 450 and 1000 nucleotide bases in length, are obtained from a total RNA pool. These amplicons are arrayed on a nylon membrane or other appropriate microchip substrate, which is then used as a regulatory gene-specific mincroarray that is hybridized with sample. Sample will typically be the mRNA obtained from cells associated with a particular state (examples include age or exposure to conditions such as outspace, low gravity), disease (such as cancer or an infection), or disorder (such as a genetic defect or trauma). The transcriptionally regulated profile of regulatory generelated genes specific to a given cultured cell sample is then determined using a software based analysis of the amount of hybridization which is detected. This information is useful in determining drug targets, markers associated with the disease state (either the presence or absence, or the extent of the disease), or the response of the disease state to drugs or other treatments.
机译:微阵列技术允许对在固体基质上从大量基因座矩阵产生的信息进行多重并行处理,这对于收集定义特定生物学状态的基因签名很有用。已经开发出一种促进该过程的方法,其中选择了相同调节方式的基因。这些基因的转录调控与相同的控制元件有关。使用已知的基因数据库,根据与其他基因的最小交叉反应性,选择特定于调控基因的引物。与该序列结合或表达取决于该结合的已知基因的选定区域的PCR产物,以及与可调节基因和控制基因相互作用的基因,称为“扩增子”或“基因cDNA片段”,介于从总RNA库获得长度为450和1000个核苷酸的碱基。将这些扩增子排列在尼龙膜或其他合适的微芯片基板上,然后将其用作与样品杂交的调控基因特异性微阵列。样品通常是从与特定状态(例如年龄或暴露于诸如空间,低重力的条件下),疾病(例如癌症或感染)或疾病(例如遗传缺陷或创伤)相关的细胞中获得的mRNA。 )。然后使用对检测到的杂交量进行基于软件的分析,确定特定于给定培养细胞样品的调控基因相关基因的转录调控特性。该信息可用于确定药物靶标,与疾病状态(疾病存在或不存在或疾病程度)相关的标记,或疾病状态对药物或其他治疗的反应。

著录项

  • 公开/公告号WO0175162A3

    专利类型

  • 公开/公告日2003-01-16

    原文格式PDF

  • 申请/专利号WO2001US10096

  • 发明设计人 WANG EUGENIA;

    申请日2001-03-29

  • 分类号C12Q1/68;

  • 国家 WO

  • 入库时间 2022-08-21 23:56:12

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