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    Use of VSV-G pseudotyped vectors for transfer of genes into embryos
    2.
    发明授权
    Use of VSV-G pseudotyped vectors for transfer of genes into embryos 失效
    使用VSV-G假型载体将基因转移到胚胎中

    公开(公告)号：US5670354A

    公开(公告)日：1997-09-23

    申请号：US480453

    申请日：1995-06-07

    申请人： Jane C. Burns ,  Jiing-Kuan Yee ,  Theodore Friedmann

    发明人： Jane C. Burns ,  Jiing-Kuan Yee ,  Theodore Friedmann

    IPC分类号： C12N5/00 ,  A61K38/00 ,  A61K48/00 ,  C07K14/145 ,  C12N5/10 ,  C12N7/00 ,  C12N7/04 ,  C12N15/09 ,  C12N15/867 ,  C12N15/87 ,  C12R1/91 ,  C12N15/00 ,  C12N5/06 ,  C12N5/16 ,  C12N15/06

    CPC分类号： C12N15/86 ,  A61K48/00 ,  C07K14/005 ,  C12N15/87 ,  C12N7/00 ,  A61K38/00 ,  C12N2740/13043 ,  C12N2740/13052 ,  C12N2740/13062 ,  C12N2760/20222 ,  C12N2830/002 ,  C12N2830/42 ,  C12N2830/60 ,  C12N2840/20 ,  C12N2840/203

    摘要： The present application discloses retrovirus-derived vectors in which the retroviral envelope glycoprotein has been replaced by the G glycoprotein of vesicular stomatitis virus, and the use of these vectors in the transfer of exogenous genes into the cells of a wide variety of non-mammalian organisms. Also disclosed is a method for the generation of retroviral vectors in high titers, wherein a recombinant, stable host cell line is provided which harbors the retroviral vector of interest without envelope protein. High-titer retroviral vector production is initiated by introducing nucleic acid encoding a functional membrane-associated protein into the cell line. The vectors disclosed in the present application can be concentrated by ultracentrifugation to titers greater than 10.sup.9 cfu/ml which are especially useful in human gene therapy trials, and can also infect cells, such as hamster and fish cells, that are ordinarily resistant to infection with vectors containing the retroviral envelope protein.

    摘要翻译： 本申请公开了逆转录病毒衍生载体，其中逆转录病毒包膜糖蛋白已经被水泡性口炎病毒的G糖蛋白替代，以及这些载体在将外源基因转移到多种非哺乳动物生物体的细胞中的用途 。 还公开了用于产生高滴度的逆转录病毒载体的方法，其中提供重组的稳定的宿主细胞系，其含有没有包膜蛋白的感兴趣的逆转录病毒载体。 通过将编码功能性膜相关蛋白的核酸引入细胞系来启动高效价逆转录病毒载体产生。 本申请中公开的载体可以通过超速离心浓缩至大于109cfu / ml的滴度，这在人类基因治疗试验中特别有用，并且还可以感染通常抗感染的细胞，例如仓鼠和鱼细胞 含有逆转录病毒包膜蛋白的载体。

    Inducible expression system
    3.
    发明授权
    Inducible expression system 失效
    诱导表达系统

    公开(公告)号：US06432705B1

    公开(公告)日：2002-08-13

    申请号：US09566660

    申请日：2000-05-08

    申请人： Jiing-Kuan Yee ,  Theodore Friedmann ,  Shin-Tai Chen

    发明人： Jiing-Kuan Yee ,  Theodore Friedmann ,  Shin-Tai Chen

    IPC分类号： C12N500

    CPC分类号： C12N15/86 ,  C12N2740/13043 ,  C12N2830/002 ,  C12N2830/006 ,  C12N2840/20 ,  C12N2840/203

    摘要： The present invention features compositions and methods for the inducible expression of a polypeptide, especially a polypeptide normally cytotoxic to the eukaryotic host cell in which it is to be expressed. A nucleotide sequence encoding a polypeptide of interest is operably linked to an inducible promoter. Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription, a transcriptional activation domain, and a second ligand-binding domain that positively regulates the transcriptional activation function of the transactivator. Transcription of the nucleotide sequence under control of the inducible promoter is activated by the multi-chimeric transactivator when both the ligand that binds the first ligand-binding domain is absent and the ligand that binds the second ligand-binding domain is present. This inducible expression system is particularly useful in the expression of the cytotoxic protein VSV G for the production of pseudotyped retroviral vectors.

    摘要翻译： 本发明的特征在于可诱导表达多肽的组合物和方法，特别是对待其表达的真核宿主细胞通常具有细胞毒性的多肽。 编码目的多肽的核苷酸序列与诱导型启动子可操作地连接。 来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节转录的第一配体结合结构域，转录激活结构域和正调节反式激活因子的转录激活功能的第二配体结合结构域组成 。 当两个结合第一配体结合结构域的配体和不存在结合第二配体结合结构域的配体时，多重嵌合反式激活剂可激活在诱导型启动子控制下的核苷酸序列的转录。 这种诱导表达系统特别可用于细胞毒素蛋白VSV G的表达以产生假型逆转录病毒载体。

    Inducible expression system
    5.
    发明授权
    Inducible expression system 失效
    诱导表达系统

    公开(公告)号：US6133027A

    公开(公告)日：2000-10-17

    申请号：US693940

    申请日：1996-08-07

    申请人： Jiing-Kuan Yee ,  Theodore Friedmann ,  Shin-Tai Chen

    发明人： Jiing-Kuan Yee ,  Theodore Friedmann ,  Shin-Tai Chen

    IPC分类号： C12N15/867 ,  C12N15/85 ,  C12N15/86

    CPC分类号： C12N15/86 ,  C12N2740/13043 ,  C12N2830/002 ,  C12N2830/006 ,  C12N2840/20 ,  C12N2840/203

    摘要： The present invention features compositions and methods for the inducible expression of a polypeptide, especially a polypeptide normally cytotoxic to the eukaryotic host cell in which it is to be expressed. A nucleotide sequence encoding a polypeptide of interest is operably linked to an inducible promoter (e.g, a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription (e.g., a prokaryotic tetracycline repressor polypeptide), a transcriptional activation domain, and a second ligand-binding domain that positively regulates the transcriptional activation function of the transactivator (e.g., a ligand-binding domain of a steroid receptor, preferably an estrogen receptor (ER)). Transcription of the nucleotide sequence under control of the inducible promoter is activated by the multi-chimeric transactivator when both the ligand that binds the first ligand-binding domain (e.g., tetracycline) is absent and the ligand that binds the second ligand-binding domain (e.g., a steroid) is present. This inducible expression system is particularly useful in the expression of the cytotoxic protein VSV G for the production of pseudotyped retroviral vectors.

    摘要翻译： 本发明的特征在于可诱导表达多肽的组合物和方法，特别是对待其表达的真核宿主细胞通常具有细胞毒性的多肽。 编码感兴趣多肽的核苷酸序列可操作地连接到诱导型启动子（例如，由连接至多拷贝tetO的最小启动子，大肠杆菌四环素抗性操纵子Tn10的四环素阻遏物（tetR）的结合位点， ）。 来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节转录的第一配体结合结构域（例如原核四环素阻遏物多肽），转录激活结构域和第二配体结合结构域组成 正调节反式激活因子的转录激活功能（例如，类固醇受体的配体结合结构域，优选雌激素受体（ER））。 当结合第一配体结合结构域（例如四环素）的配体和结合第二配体结合结构域的配体两者都可以通过多嵌合反式激活因子激活受诱导型启动子控制下的核苷酸序列的转录（ 例如，类固醇）存在。 这种诱导表达系统特别可用于细胞毒素蛋白VSV G的表达以产生假型逆转录病毒载体。

    Packaging cell lines for pseudotyped retroviral vectors
    7.
    发明授权
    Packaging cell lines for pseudotyped retroviral vectors 失效
    包装用于假型逆转录病毒载体的细胞系

    公开(公告)号：US5739018A

    公开(公告)日：1998-04-14

    申请号：US694652

    申请日：1996-08-07

    申请人： Atsushi Miyanohara ,  Jiing-Kuan Yee ,  Shin-Tai Chen ,  Charles Edward Prussak ,  Theodore Friedmann

    发明人： Atsushi Miyanohara ,  Jiing-Kuan Yee ,  Shin-Tai Chen ,  Charles Edward Prussak ,  Theodore Friedmann

    IPC分类号： C12N15/63 ,  C12N15/867 ,  C12N5/10 ,  C12N15/86

    CPC分类号： C12N15/86 ,  C12N15/635 ,  C12N2740/13052 ,  C12N2830/002 ,  C12N2830/006 ,  C12N2830/15 ,  C12N2830/30 ,  C12N2840/20 ,  C12N2840/203

    摘要： The present invention features packaging cell lines and recombinant retroviral particles produced thereby, particularly pseudotyped retroviral particles. Preferably, the packaging cell lines are derived from HeLa, Cf2Th, D17, MDCK, or BHK cells, most preferably from Cf2Th cells. Retroviral particles are produced by inducibly expressing an envelope protein of interest (e.g., a retroviral envelope or the envelope protein of vesicular stomatitis virus (VSV G)). Inducible expression of the envelope protein is accomplished by operably linking an envelope protein-encoding nucleotide sequence to an inducible promoter (e.g., a promoter composed of a minimal promoter linked to multiple copies of tetO, the binding site for the tetracycline repressor (tetR) of the Escherichia coli, tetracycline resistance operon Tn10). Expression from the inducible promoter is regulated by a multi-chimeric transactivating factor, composed of a first ligand-binding domain that negatively regulates transcription from the inducible promoter (e.g., a prokaryotic tetracycline repressor polypeptide (tetR)), a transcriptional activation domain, and a second ligand-binding domain (e.g., a ligand-binding domain of a steroid receptor, preferably an estrogen receptor (ER)).

    摘要翻译： 本发明的特征在于包装细胞系和由此产生的重组逆转录病毒颗粒，特别是假型逆转录病毒颗粒。 优选地，包装细胞系衍生自HeLa，Cf2Th，D17，MDCK或BHK细胞，最优选来自Cf2Th细胞。 逆转录病毒颗粒通过诱导表达感兴趣的包膜蛋白（例如，逆转录病毒包膜或水泡性口炎病毒的包膜蛋白（VSV G））产生。 通过将包膜蛋白编码核苷酸序列可操作地连接到诱导型启动子（例如，由与tetO的多拷贝连接的最小启动子，四环素阻遏物（tetR）的结合位点）组成的启动子来实现包膜蛋白的诱导表达 大肠杆菌，四环素抗性操纵子Tn10）。 来自诱导型启动子的表达由多嵌合反式激活因子调节，所述多嵌合反式激活因子由负调节来自诱导型启动子的转录的第一配体结合结构域（例如，原核四环素阻遏物多肽（tetR）），转录激活结构域和 第二配体结合结构域（例如，类固醇受体的配体结合结构域，优选雌激素受体（ER））。

    Method for retrovirus vector production by separated gag and pol expression
    9.
    发明授权
    Method for retrovirus vector production by separated gag and pol expression 失效
    通过分离的gag和pol表达产生逆转录病毒载体的方法

    公开(公告)号：US06451304B1

    公开(公告)日：2002-09-17

    申请号：US09265013

    申请日：1999-03-09

    申请人： Theodore Friedmann ,  Atsushi Miyanohara

    发明人： Theodore Friedmann ,  Atsushi Miyanohara

    IPC分类号： A01N6300

    CPC分类号： C12N15/86 ,  C12N2740/13043 ,  C12N2740/16043

    摘要： The invention provides a system for production of retroviruses which are replication incompetent. In the system, gag and pol retroviral structural proteins are expressed separately by different plasmids in a packaging cell line. Separate gag and pol expressing plasmids are provided, as are packaging cell lines containing such plasmids. Retrovirus products of the retroviral vector production system, including chimeric retroviruses, are also provided.

    摘要翻译： 本发明提供了一种用于生产复制无能力的逆转录病毒的系统。 在系统中，gag和pol逆转录病毒结构蛋白在包装细胞系中由不同质粒分开表达。 提供单独的gag和pol表达质粒，以及含有这种质粒的包装细胞系也是如此。 还提供了逆转录病毒载体生产系统的逆转录病毒产品，包括嵌合逆转录病毒。