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    • 8. 发明授权
    • Tetracycline-inducible transcriptional inhibitor fusion proteins
    • 四环素诱导型转录抑制剂融合蛋白
    • US06271348B1
    • 2001-08-07
    • US09489777
    • 2000-01-24
    • Hermann BujardManfred Gossen
    • Hermann BujardManfred Gossen
    • C07K1900
    • C07K14/005A01K67/0275A01K2217/05A01K2217/075A01K2217/20A01K2227/105A01K2267/0393A61K48/00C07K14/245C07K2319/00C07K2319/09C07K2319/61C07K2319/71C07K2319/80C12N15/62C12N15/63C12N15/635C12N15/67C12N15/85C12N15/8509C12N2710/16622C12N2830/003C12N2830/006C12N2830/32
    • Methods of regulating gene expression in subjects using tetracycline-responsive fusion proteins are disclosed. In one embodiment, the method involves introducing into a cell the subject a nucleic acid molecule encoding a fusion protein which inhibits transcription, the fusion protein comprising a first polypeptide which binds to a tet operator sequence, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first polypeptide can binds to a tet operator sequence in the absence, but not the presence, of tetracycline. Alternatively, the first polypeptide can binds to a tet operator sequence in the presence, but not the absence, of tetracycline. In another embodiment, the method of the invention involves obtaining a cell from a subject, introducing into the cell a first nucleic acid molecule which operatively links a gene to at least one tet operator sequence, introducing into the cell a second nucleic acid molecule encoding an inhibitory fusion protein of the invention to form a modified cell, administering the modified cell to the subject and modulating the concentration of a tetracycline, or analogue thereof, in the subject. The first and second nucleic acid molecules can be linked or can be separate molecules.
    • 公开了使用四环素响应融合蛋白调节受试者基因表达的方法。 在一个实施方案中,该方法包括向受试者引入编码抑制转录的融合蛋白的核酸分子,该融合蛋白包含与tet操纵子序列结合的第一多肽,该第一多肽与抑制转录的异源第二多肽有效连接 在真核细胞中 并调节受试者中四环素或其类似物的浓度。 在不存在但不存在四环素的情况下,第一种多肽可以结合tet操纵子序列。 或者,第一多肽可以在四环素存在下但不存在时与tet操纵子序列结合。 在另一个实施方案中,本发明的方法涉及从受试者获得细胞,将在第一核酸分子中引入至少一个tet操纵子序列的第一核酸分子,将编码 本发明的抑制性融合蛋白以形成修饰的细胞,向受试者施用修饰的细胞并调节受试者中四环素或其类似物的浓度。 第一和第二核酸分子可以连接或可以是分开的分子。