Methods for making polynucleotide libraries, polynucleotide arrays, and cell libraries for high-throughput genomics analysis
    70.
    发明申请
    Methods for making polynucleotide libraries, polynucleotide arrays, and cell libraries for high-throughput genomics analysis 审中-公开
    用于制备用于高通量基因组学分析的多核苷酸文库,多核苷酸阵列和细胞文库的方法

    公开(公告)号:US20020150945A1

    公开(公告)日:2002-10-17

    申请号:US10172715

    申请日:2002-06-13

    CPC classification number: C12N15/85 C12N15/10 C12N15/1034 C12N15/902

    Abstract: A method for high-throughput genomics analysis, to identify the therapeutic or diagnostic utility of genes, entails the use of a construct to disrupt a gene or alleles of a gene in cells of interest. Arrays of such cells can be used to monitor such disrupted cells phenotypically in the context, for example, of testing drug candidates. Polynucleotides that comprise part of the disrupted genes can be recovered from such nullknockoutnull cells, by virtue of an origin of replication or a host cell selection marker sequence that is part of the construct. The recovered polynucleotides can be used to identify the disrupted genes or to make homologous recombination vectors, which in turn can be employed to make multi-allele knockout cells. Double-stranded RNA molecules designed to target the recovered polynucleotide are used to downregulate the polynucleotide in vitro and in vivo, following determination of a therapeutically effective dosage of the RNAi molecule.

    Abstract translation: 用于识别基因的治疗或诊断效用的高通量基因组学分析的方法需要使用构建体来破坏目的细胞中基因的基因或等位基因。 这种细胞的阵列可以用于在上下文中表型地监测这样的破坏细胞,例如测试候选药物。 可以通过复制起点或作为构建体一部分的宿主细胞选择标记序列从这种“敲除”细胞中回收构成部分破坏基因的多核苷酸。 回收的多核苷酸可用于鉴定破坏的基因或使同源重组载体,其又可用于制备多等位基因敲除细胞。 在测定治疗有效剂量的RNAi分子后,设计用于靶向回收多核苷酸的双链RNA分子用于体外和体内下调多核苷酸。

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