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    TETRACYCLINE INDUCIBLE TRANSCRIPTION CONTROL SEQUENCE
    1.
    发明申请
    TETRACYCLINE INDUCIBLE TRANSCRIPTION CONTROL SEQUENCE 有权
    四环素诱导转录控制序列

    公开(公告)号：US20110247088A1

    公开(公告)日：2011-10-06

    申请号：US13121673

    申请日：2009-08-19

    申请人： Hermann Bujard ,  Rainer Loew

    发明人： Hermann Bujard ,  Rainer Loew

    IPC分类号： A01K67/00 ,  C12N15/63 ,  C12N5/10 ,  C12N1/00 ,  C07H21/04 ,  A01H5/00 ,  C12N15/82

    CPC分类号： C12N15/635 ,  C12N2770/40043 ,  C12N2830/003

    摘要： The present invention relates to inducible transcription control sequences for the regulation of gene expression. Specifically, it relates to an transcription control sequence comprising at least two tet operator sequence motifs allowing the binding of tetracycline-dependent transcriptional regulators, wherein each of the said tetracycline-dependent transcriptional regulators binds with respect to its neighbour to an opposite face of the DNA helix, and a minimal promoter comprising a TATA box which is linked at its 5′ end to a general transcription factor binding site. Further, the present invention relates to a vector, a host cell, a plant or a non-human transgenic animal comprising the said transcription control sequence. Also contemplated is a method for regulating the expression of a nucleic acid sequence being operatively linked to the transcription control sequence of the present invention in a host cell, a plant or a non-human transgenic animal.

    摘要翻译： 本发明涉及用于调节基因表达的诱导型转录调控序列。 具体地说，本发明涉及包含至少两个允许四环素依赖性转录调节子结合的tet操纵子序列基序的转录控制序列，其中所述四环素依赖性转录调节子中的每一个相对于DNA的相对面与DNA的相邻结合 螺旋和包含在其5'末端与一般转录因子结合位点连接的TATA盒的最小启动子。 此外，本发明涉及包含所述转录调控序列的载体，宿主细胞，植物或非人转基因动物。 还考虑了在宿主细胞，植物或非人转基因动物中调节与本发明的转录调控序列有效连接的核酸序列的表达的方法。

    Tet repressor-based transcriptional regulatory proteins
    2.
    发明授权
    Tet repressor-based transcriptional regulatory proteins 有权
    基于阻遏物的转录调控蛋白

    公开(公告)号：US07541446B2

    公开(公告)日：2009-06-02

    申请号：US10456395

    申请日：2003-06-05

    申请人： Wolfgang Hillen ,  Hermann Bujard ,  Stefanie Urlinger

    发明人： Wolfgang Hillen ,  Hermann Bujard ,  Stefanie Urlinger

    IPC分类号： C07H21/04 ,  C12N15/63

    CPC分类号： C12N15/635 ,  C07K2319/00 ,  C12N15/85 ,  C12N2800/108 ,  C12N2830/003 ,  C12N2830/006

    摘要： The present invention provides a panel of transcriptional activator fusion proteins which comprises both tetracycline controlled transactivator proteins and reverse tetracycline transactivator proteins. These transactivators have novel phenotypes such as altered basal transcriptional activity in the absence of doxycycline, altered induced transcriptional activity in the presence of doxycycline, or differential induction by tetracycline and analogs of tetracycline.

    摘要翻译： 本发明提供了一组转录激活物融合蛋白，其包含四环素控制的反式激活蛋白和反向四环素反式激活蛋白。 这些反式激活剂具有新型表型，例如在多西环素不存在时改变的基础转录活性，在多西环素存在下改变的诱导转录活性，或四环素和四环素类似物的差异诱导。

    Methods for regulating gene expression
    4.
    发明授权
    Methods for regulating gene expression 失效
    调节基因表达的方法

    公开(公告)号：US06783756B2

    公开(公告)日：2004-08-31

    申请号：US09281674

    申请日：1999-03-30

    申请人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

    发明人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

    IPC分类号： A61K6300

    CPC分类号： C12N15/62 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N15/635 ,  C12N15/85 ,  C12N2830/006

    摘要： A method for regulating expression of a tet operator-linked gene in a cell of a subject is disclosed. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a tetracycline-controllable transactivator (tTA), the tTA comprising a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription in eucaryotic cells; and modulating the concentration of a tetracycline, or analogue thereof, in the subject. Alternatively, in another embodiment, the method involves obtaining the cell from the 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 a tTA, 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 molecule can be within a single molecule (e.g., in the same vector) or on separate molecules.

    摘要翻译： 公开了一种用于调节受试者细胞中tet操纵子连接基因表达的方法。 在一个实施方案中，所述方法包括将编码四环素可控反式激活因子（tTA）的核酸分子引入细胞，所述tTA包含可操作地连接到直接或间接激活真核细胞中的转录的多肽的Tet阻遏物; 并调节受试者中四环素或其类似物的浓度。 或者，在另一个实施方案中，所述方法包括从受试者获得细胞，向细胞中引入第一核酸分子，该第一核酸分子可操作地将基因连接至至少一个tet操纵子序列，将编码tTA的第二核酸分子 ，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在单分子内（例如，在同一载体中）或分开的分子上。

    Transgenic organisms having tetracycline-regulated transcriptional regulatory systems
    5.
    发明授权
    Transgenic organisms having tetracycline-regulated transcriptional regulatory systems 有权
    具有四环素调节转录调控系统的转基因生物

    公开(公告)号：US06252136B1

    公开(公告)日：2001-06-26

    申请号：US09163269

    申请日：1998-09-29

    申请人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

    发明人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

    IPC分类号： C12N1509

    CPC分类号： C12N15/62 ,  A01K67/0275 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N15/635 ,  C12N15/85 ,  C12N2830/006

    摘要： Transgenic animals carrying two transgenes, the first coding for a transactivator fusion protein comprising a tet repressor and a polypeptide which directly or indirectly activates in eucaryotic cells, and the second comprising a gene operably linked to a minimal promoter operably linked to at least one tet operator sequence, are disclosed. Isolated DNA molecules (e.g., targeting vectors) for integrating a polynucleotide sequence encoding a transactivator of the invention at a predetermined location within a second target DNA molecule by homologous recombination are also disclosed. Transgenic animals having the DNA molecules of the invention integrated at a predetermined location in a chromosome by homologous recombination are also encompassed by the invention. Methods to regulate the expression of a tet operator linked-gene of interest by administering tetracycline or a tetracycline analogue to an animal of the invention are also disclosed. The regulatory system of the invention allows for conditional inactivation or modulation of expression of a gene of interest in a host cell or animal.

    摘要翻译： 携带两个转基因的转基因动物，第一编码包含直接或间接在真核细胞中活化的tet阻遏物和多肽的反式激活因子融合蛋白，第二编码包含与可操作地连接至少一个tet操纵子的最小启动子可操作地连接的基因 序列。 还公开了通过同源重组将编码本发明的反式激活物的多核苷酸序列整合到第二靶DNA分子内的预定位置的分离的DNA分子（例如靶向载体）。 通过同源重组将具有整合在染色体中预定位置的本发明的DNA分子的转基因动物也包括在本发明中。 还公开了通过向本发明的动物施用四环素或四环素类似物来调节所述目的基因的表达的方法。 本发明的调节系统允许在宿主细胞或动物中有条件地失活或调控感兴趣的基因的表达。

    Methods for regulating gene expression
    7.
    发明授权
    Methods for regulating gene expression 失效
    调节基因表达的方法

    公开(公告)号：US5814618A

    公开(公告)日：1998-09-29

    申请号：US485978

    申请日：1995-06-07

    申请人： Hermann Bujard ,  Manfred Gossen

    发明人： Hermann Bujard ,  Manfred Gossen

    IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C07K14/035 ,  C07K14/195 ,  C07K14/245 ,  C07K19/00 ,  C12N15/11 ,  C12N15/63 ,  C12N15/67 ,  C12N15/85 ,  C12N15/00

    CPC分类号： C07K14/005 ,  A01K67/0275 ,  C07K14/245 ,  C12N15/62 ,  C12N15/63 ,  C12N15/635 ,  C12N15/67 ,  C12N15/85 ,  C12N15/8509 ,  A01K2217/05 ,  A01K2217/075 ,  A01K2217/20 ,  A01K2227/105 ,  A01K2267/0393 ,  A61K48/00 ,  C07K2319/00 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2710/16622 ,  C12N2830/003 ,  C12N2830/006 ,  C12N2830/32

    摘要： 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.

    摘要翻译： 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。

    Tight control of gene expression in eucaryotic cells by tetracycline-responsive promoters
    8.
    发明授权
    Tight control of gene expression in eucaryotic cells by tetracycline-responsive promoters 失效
    通过四环素响应启动子对真核细胞中基因表达的严格控制

    公开(公告)号：US5650298A

    公开(公告)日：1997-07-22

    申请号：US260452

    申请日：1994-06-14

    申请人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

    发明人： Hermann Bujard ,  Manfred Gossen ,  Jochen G. Salfeld ,  Jeffrey W. Voss

    IPC分类号： A01K67/027 ,  A61K48/00 ,  C07K14/00 ,  C12N5/08 ,  C12N5/10 ,  C12N15/00 ,  C12N15/09 ,  C12N15/31 ,  C12N15/62 ,  C12N15/63 ,  C12N15/85 ,  C12R1/91 ,  C12P21/00 ,  C07H21/04

    CPC分类号： A01K67/0275 ,  C12N15/62 ,  C12N15/635 ,  C12N15/85 ,  A01K2217/05 ,  A01K2217/075 ,  C07K2319/09 ,  C07K2319/61 ,  C07K2319/71 ,  C07K2319/80 ,  C12N2830/006

    摘要： Transgenic animals carrying two transgenes, the first coding for a transactivator fusion protein comprising a tet repressor and a polypeptide which directly or indirectly activates in eucaryotic cells, and the second comprising a gene operably linked to a minimal promoter operably linked to at least one tet operator sequence, are disclosed. Isolated DNA molecules (e.g., targeting vectors) for integrating a polynucleotide sequence encoding a transactivator of the invention at a predetermined location within a second target DNA molecule by homologous recombination are also disclosed. Transgenic animals having the DNA molecules of the invention integrated at a predetermined location in a chromosome by homologous recombination are also encompassed by the invention. Methods to regulate the expression of a tet operator linked-gene of interest by administering tetracycline or a tetracycline analogue to an animal of the invention are also disclosed. The regulatory system of the invention allows for conditional inactivation or modulation of expression of a gene of interest in a host cell or animal.

    摘要翻译： 携带两个转基因的转基因动物，第一编码包含直接或间接在真核细胞中活化的tet阻遏物和多肽的反式激活因子融合蛋白，第二编码包含与可操作地连接至少一个tet操纵子的最小启动子可操作地连接的基因 序列。 还公开了通过同源重组将编码本发明的反式激活物的多核苷酸序列整合到第二靶DNA分子内的预定位置的分离的DNA分子（例如靶向载体）。 通过同源重组将具有整合在染色体中预定位置的本发明的DNA分子的转基因动物也包括在本发明中。 还公开了通过向本发明的动物施用四环素或四环素类似物来调节所述目的基因的表达的方法。 本发明的调节系统允许在宿主细胞或动物中有条件地失活或调控感兴趣的基因的表达。

    Transcriptional activators with graded transactivation potential
    10.
    发明授权
    Transcriptional activators with graded transactivation potential 有权
    具有分级反式激活潜能的转录激活子

    公开(公告)号：US06271341B1

    公开(公告)日：2001-08-07

    申请号：US09577027

    申请日：2000-05-23

    申请人： Udo Baron ,  Manfred Gossen ,  Hermann Bujard

    发明人： Udo Baron ,  Manfred Gossen ,  Hermann Bujard

    IPC分类号： C07K1900

    CPC分类号： C07K14/005 ,  A01K2217/05 ,  A61K48/00 ,  C07K2319/00 ,  C12N15/635 ,  C12N15/85 ,  C12N2710/16622 ,  C12N2830/003 ,  C12N2830/006

    摘要： Transcriptional activators which differ in their activation potential by more than 3 orders of magnitude are provided. The transactivators are fusions between a DNA binding protein (e.g., a Tet repressor) and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). Substitution mutations at amino acid position 442 within the minimal VP16 domain provide transactivators with differing transactivation ability. Moreover, chimeric activation domains comprising both wild type and mutant minimal VP16 domains provide additional variants with differing transactivation ability. Various aspects of the invention pertain to nucleic acid molecules, vectors, host cells, fusion proteins, transgenic and homologous recombinant organisms and methods of regulating gene transcription.

    摘要翻译： 提供了其激活电位不同于3个数量级的转录激活子。 反式激活因子是DNA结合蛋白（例如Tet阻遏物）和衍生自单纯疱疹病毒蛋白16（VP16）的最小转录激活结构域之间的融合物。 在最小VP16结构域内的氨基酸位置442处的取代突变体提供具有不同反式激活能力的反式激活因子。 此外，包含野生型和突变型最小VP16结构域的嵌合活化域提供了具有不同反式激活能力的另外的变体。 本发明的各个方面涉及核酸分子，载体，宿主细胞，融合蛋白，转基因和同源重组生物以及调节基因转录的方法。