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    Transfection of vertebrate cells e.g. by homologous recombination
    7.
    发明公开
    Transfection of vertebrate cells e.g. by homologous recombination 失效
    转基因脊椎动物疫苗组合

    公开(公告)号：EP1221477A2

    公开(公告)日：2002-07-10

    申请号：EP01204619.9

    申请日：1992-11-05

    申请人： TRANSKARYOTIC THERAPIES, INC.

    发明人： Selden, Richard F. ,  Heartlein, Michael W. ,  Treco, Douglas A.

    IPC分类号： C12N5/10 ,  C12N15/85 ,  A61K48/00

    CPC分类号： C12N15/907 ,  A61K47/6901 ,  A61K48/00 ,  C07K14/505 ,  C07K14/605 ,  C07K14/61 ,  C07K2319/02 ,  C12N15/85 ,  C12N2510/02 ,  C12N2800/108 ,  C12N2830/002 ,  C12N2830/85 ,  C12N2840/20 ,  C12N2840/44

    摘要： The present invention relates to transfected primary and secondary somatic cells of vertebrate origin, particularly mammalian origin transfected with exogenous genetic material (DNA) which encodes a desired (e.g. a therapeutic) product or is itself a desired (e.g. therapeutic) product, methods by which primary and secondary cells are transfected to include exogenous genetic material, including DNA targeting by homologous recombination, methods of producing clonal cell strains or heterogenous cell strains, methods of gene therapy in which the transfected primary or secondary cells are used, and methods of producing antibodies using the transfected primary or secondary cells. The present invention includes primary and secondary somatic cells, such as fibroblasts, keratinocytes, epithelial cells, endothelial cells, glial cells, neural cells, formed elements of the blood, muscle cells, other somatic cells which can be cultured and somatic cell precursors, which have been transfected with exogenous DNA which is stably integrated into their genomes or is expressed in the cells episomally. The exogenous DNA either encodes a product, such as a translational product (e.g. a protein) or a transcriptional product (e.g. a ribozyme or an anti-sense nucleic acid sequence) which is a therapeutic product or is itself a therapeutic product (e.g. DNA which binds to a cellular regulatory protein or alters gene expression).

    摘要翻译： 本发明涉及脊椎动物来源的转染的初级和次级体细胞，特别是用编码期望（例如治疗性）产品的外源性遗传物质（DNA）转染的哺乳动物来源，或本身是所需（例如治疗性）产品的方法， 转染原代细胞和二次细胞以包括外源遗传物质，包括通过同源重组进行DNA靶向，产生克隆细胞株或异源细胞株的方法，使用转染的原代细胞或次细胞的基因治疗方法，以及产生抗体的方法 使用转染的一次或二次细胞。 本发明包括初级和次级体细胞，例如成纤维细胞，角质形成细胞，上皮细胞，内皮细胞，神经胶质细胞，神经细胞，形成的血液元素，肌肉细胞，可培养的其他体细胞和体细胞前体，其中 已经用稳定整合到其基因组中的外源DNA转染，或者以细胞的形式表达。 外源DNA编码产物，例如翻译产物（例如蛋白质）或作为治疗产物的转录产物（例如核酶或反义核酸序列），或本身是治疗产物（例如DNA 结合细胞调节蛋白或改变基因表达）。

    ACTIVATING EXPRESSION OF AN AMPLIFYING ENDOGENOUS GENE BY HOMOLOGOUS RECOMBINATION
    8.
    发明公开
    ACTIVATING EXPRESSION OF AN AMPLIFYING ENDOGENOUS GENE BY HOMOLOGOUS RECOMBINATION 失效
    EXPRIMIERUNGSAKTIVATION和内源性GEN再现同源重组。

    公开(公告)号：EP0672160A1

    公开(公告)日：1995-09-20

    申请号：EP94903405.0

    申请日：1993-12-02

    申请人： TRANSKARYOTIC THERAPIES, INC.

    发明人： TRECO, Douglas, A. ,  HEARTLEIN, Michael, W. ,  SELDEN, Richard, F.

    IPC分类号： C12N15 ,  A61K31 ,  A61K35 ,  A61K38 ,  A61K48 ,  C07H21 ,  C07K14 ,  C12N5

    CPC分类号： C12N15/90 ,  A61K38/00 ,  A61K48/00 ,  C07K14/505 ,  C07K14/61 ,  C07K2319/036 ,  C07K2319/75 ,  C12N15/907

    摘要： Method of activating expression of and amplifying an endogenous gene in genomic DNA of a vertebrate cell which is not expressed in the cell as obtained or is not expressed at significant levels in the cell as obtained, comprising the steps of: (a) transfecting cells with DNA sequences comprising: 1) exogenous DNA sequences which repair, alter, delete or replace a sequence present in the cell or which are regulatory sequences not normally functionally linked to the endogenous gene in the cell as obtained; 2) DNA sequences homologous with genomic DNA sequences at a preselected site in the cells; and 3) amplifiable DNA encoding a selectable marker; (b) maintaining the cells under conditions appropriate for homologous recombination to occur between DNA sequences homologous with genomic DNA sequences and genomic DNA sequences; (c) culturing the homologously recombinant cells produced in (b) under conditions which select from amplification of the amplifiable DNA encoding a selectable marker, whereby the amplifiable DNA encoding a selectable marker and the endogenous gene functionally linked to exogenous DNA of (a) 1) are coamplified.