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    • 4. 发明申请
    • A Lac REPRESSOR-HSV VP16 CHIMERIC TRANSCRIPTIONAL ACTIVATOR PROTEIN SYSTEM FUNCTIONING IN TRANSFECTED MAMMALIAN CELLS
    • 转基因哺乳动物细胞中的Lac REPRESSOR-HSV VP16基因转录激活蛋白系统功能
    • WO1991019784A1
    • 1991-12-26
    • PCT/US1991004180
    • 1991-06-12
    • THE TRUSTEES OF PRINCETON UNIVERSITY
    • THE TRUSTEES OF PRINCETON UNIVERSITYBAIM, Steven, B.LABOW, Mark, A.SHENK, Thomas, E.LEVINE, Arnold, J.
    • C12N05/06
    • C12N15/85C07K14/005C12N2710/16622C12N2800/108C12N2830/00C12N2830/002C12N2830/15C12N2830/55C12N2830/60C12N2830/85C12N2840/44
    • The present invention relates to novel chimeric transactivating proteins comprising a functional portion of a DNA binding protein and a functional portion of a transcriptional activator protein. The chimeric transactivating proteins of the invention offer a variety of advantages, including the specific activation of expression of genes engineered to comprise transactivator responsive elements, thereby achieving exceptionally high levels of gene expression. Furthermore, in various embodiments of the invention, the transactivator proteins may be used to increase expression of some genes while repressing the expression of others, thus permitting a greater degree of control of gene expression patterns than other currently available systems. In preferred embodiments of the invention, the function of the chimeric lac repressor-HSVVP16 tansactivator proteins may be induced, for example, by chemical agents such as IPTG or changes in temperature. In a preferred specific embodiment of the invention, the ability to switch transactivator function on and off may be utilized in the context of transgenic animals and to develop cell lines capable of differentiation.
    • 本发明涉及包含DNA结合蛋白的功能部分和转录激活蛋白的功能部分的新型嵌合反式激活蛋白。 本发明的嵌合反式激活蛋白提供多种优点,包括工程改造成包含反式激活因子反应元件的基因的表达的特异性激活,从而获得异常高水平的基因表达。 此外,在本发明的各种实施方案中,反式激活蛋白可用于增加某些基因的表达,同时抑制其他基因的表达,从而允许比其它目前可用的系统更大程度地控制基因表达模式。 在本发明的优选实施方案中,可以例如通过化学试剂例如IPTG或温度变化来诱导嵌合lac阻遏物-HSVVP16灭活蛋白的功能。 在本发明的优选具体实施方案中,可以在转基因动物的上下文中使用切换反式激活因子功能的能力,并开发能够分化的细胞系。
    • 6. 发明申请
    • METHODS FOR DETERMINING MULTIPLE EFFECTS OF DRUGS THAT MODULATE FUNCTION OF TRANSCRIPTION REGULATORY PROTEINS
    • 确定调节功能的转录调节蛋白的药物的多种作用的方法
    • WO2003033744A1
    • 2003-04-24
    • PCT/US2002/033406
    • 2002-10-18
    • TRUSTEES OF PRINCETON UNIVERSITYNOTTERMAN, DanielSHENK, Thomas, E.
    • NOTTERMAN, DanielSHENK, Thomas, E.
    • C12Q1/68
    • C12Q1/6809C12Q2565/501
    • Assays for determining the downstream effects of drugs that modulate the function of transcription regulatory proteins are disclosed. The assays comprise the steps of (a) providing cells that contain the transcription factor; (b) maintaining a control population and a test population of the cells under conditions that allow gene expression to occur in the cells, wherein the test population is exposed to the agent that modulates the activity of the transcription factor; (c) generating a gene expression profile for each of the control population and the test population of cells; and (d) comparing the gene expression profile from the control population of cells with the gene expression profile from the test population of cells; those differences being attributable to the effect of the agent that modulates the activity of the transcription factor. The assays are useful for classifying candidate transcription factor-modulating drugs on the basis of downstream gene expression effects, the thereafter correlating observed downstream effects with physiological markers of drug efficacy or side effects.
    • 公开了测定调节转录调节蛋白功能的药物的下游作用的方法。 测定法包括以下步骤:(a)提供含有转录因子的细胞; (b)在允许在细胞中发生基因表达的条件下维持对照群体和测试群体,其中所述测试群体暴露于调节转录因子活性的试剂; (c)产生每个对照群体和细胞的测试群体的基因表达谱; 和(d)将来自细胞对照群体的基因表达谱与来自细胞的测试群体的基因表达谱进行比较; 这些差异归因于调节转录因子活性的药剂的作用。 该测定可用于基于下游基因表达效应对候选转录因子调节药物进行分类,此后将观察到的下游作用与药物功效或副作用的生理学标记相关联。