公开(公告)号：US20080064040A1

公开(公告)日：2008-03-13

申请号：US11819769

申请日：2007-06-29

申请人： Stephen Watson Michnick ,  Barbara Belisle ,  Marnie MacDonald ,  John Westwick ,  Jane Lamerdin

发明人： Stephen Watson Michnick ,  Barbara Belisle ,  Marnie MacDonald ,  John Westwick ,  Jane Lamerdin

IPC分类号： C12Q1/68 ,  G01N33/50

CPC分类号： G01N33/502 ,  C12Q1/6897 ,  G01N33/5008 ,  G01N33/5041

摘要： The cell-based assays described in the present invention can be used to directly assess the sensitivity and specificity of the gene annotation reagent against its target, and to determine if a non-targeted gene participates in a pathway of interest or is functionally linked to another gene or protein. The combination of annotation reagents with such cell-based assays is useful for mapping genes (proteins) into cellular pathways on a genome-wide scale. Preferred assay embodiments include fluorescence or luminescence assays in intact (live or fixed) cells. Such fluorescence or luminescence assays include high-throughput or high-content assays for protein activity, subcellular localization, post-translational modifications, or interactions of proteins. Suitable assays may include protein-protein interaction assays; protein translocation assays; and post-translational modification assays. The invention can be used to assess the efficacy of any gene silencing experiment, to determine the level of gene silencing that is achieved, and to map novel genes into biochemical pathways, and to identify novel pharmaceutical targets. The results also demonstrate the feasibility of employing this strategy in genome-wide functional annotation efforts.

摘要翻译： 本发明中描述的基于细胞的测定可用于直接评估基因注释试剂对其靶标的敏感性和特异性，并确定非靶向基因是否参与感兴趣的途径或与另一种功能相关 基因或蛋白质。 注释试剂与这种基于细胞的测定法的组合可用于将基因（蛋白质）在基因组范围内映射到细胞途径中。 优选的测定实施方案包括在完整（活的或固定的）细胞中的荧光或发光测定。 这种荧光或发光测定法包括用于蛋白质活性，亚细胞定位，翻译后修饰或蛋白质相互作用的高通量或高含量测定。 合适的测定可以包括蛋白质 - 蛋白质相互作用测定; 蛋白质易位测定; 和翻译后修饰测定。 本发明可用于评估任何基因沉默实验的功效，确定实现的基因沉默水平，并将新基因映射到生物化学途径中，并鉴定新的药物靶标。 结果还表明在全基因组功能注释方面采用这一策略的可行性。

公开(公告)号：US20120208197A1

公开(公告)日：2012-08-16

申请号：US13137605

申请日：2011-08-29

申请人： Stephen William Watson Michnick ,  Barbara Belisle ,  Marnie L. MacDonald ,  John K. Westwick ,  Jane Elizabeth Lamerdin

发明人： Stephen William Watson Michnick ,  Barbara Belisle ,  Marnie L. MacDonald ,  John K. Westwick ,  Jane Elizabeth Lamerdin

IPC分类号： C12Q1/68 ,  G01N33/573 ,  G01N33/566

CPC分类号： G01N33/502 ,  C12Q1/6897 ,  G01N33/5008 ,  G01N33/5041

摘要： The cell-based assays described in the present invention can be used to directly assess the sensitivity and specificity of the gene annotation reagent against its target, mapping genes and to determine if a non-targeted gene participates in a pathway of interest or is functionally linked to another gene or protein. Preferred assay embodiments include fluorescence or luminescence assays in intact (live or fixed) cells. Such fluorescence or luminescence assays include high-throughput or high-content assays for protein activity, subcellular localization, post-translational modifications, or interactions of proteins. Suitable assays may include protein-protein interaction assays; protein translocation assays; and post-translational modification assays. The invention can be used to assess the efficacy of any gene silencing experiment, and to map novel genes into biochemical pathways, and to identify novel pharmaceutical targets. The results also demonstrate the feasibility of employing this strategy in genome-wide functional annotation efforts.

摘要翻译： 本发明中描述的基于细胞的测定可用于直接评估基因注释试剂对其靶标，测绘基因的灵敏度和特异性，并确定非靶向基因是否参与感兴趣的途径或功能上相关 到另一种基因或蛋白质。 优选的测定实施方案包括在完整（活的或固定的）细胞中的荧光或发光测定。 这种荧光或发光测定法包括用于蛋白质活性，亚细胞定位，翻译后修饰或蛋白质相互作用的高通量或高含量测定。 合适的测定可以包括蛋白质 - 蛋白质相互作用测定; 蛋白质易位测定; 和翻译后修饰测定。 本发明可用于评估任何基因沉默实验的功效，并将新基因映射到生物化学途径中，并鉴定新的药物靶点。 结果还表明在全基因组功能注释方面采用这一策略的可行性。