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    METHODS FOR HOMOLOGOUS RECOMBINATION
    12.
    发明公开
    METHODS FOR HOMOLOGOUS RECOMBINATION 有权
    方法进行同源重组

    公开(公告)号：EP1263972A2

    公开(公告)日：2002-12-11

    申请号：EP01918578.4

    申请日：2001-03-13

    申请人： TRANSKARYOTIC THERAPIES, INC.

    发明人： IVANOV, Evguenii

    IPC分类号： C12N15/63

    CPC分类号： C12N15/907 ,  C07K14/4712 ,  C12N15/63

    摘要： The invention features a method of promoting an alteration at a selected site in a target DNA, e.g., in the chromosomal DNA of a cell. The method includes providing, at the site: (a) a double stranded DNA sequence which includes a selected DNA sequence; (b) an agent which enhances homologous recombination, e.g., a Rad52 protein or a functional fragment thereof; and (c) an agent which inhibits non-homologous end joining, e.g., an agent which inactivates Ku such as an anti-Ku antibody or a Ku-binding oligomer or polymer, and allowing the alteration to occur. The agent which inhibits non-homologous end joining, e.g., a Ku inactivating agent such as an anti-Ku antibody, is preferably provided locally. Component (a), (b), and (c) can be introduced together, which is preferred, or separately.

    DNA constructs and transfection of cells therewith for homologous recombination
    15.
    发明公开
    DNA constructs and transfection of cells therewith for homologous recombination 失效
    DNA-Konstruktionen und Transfection von Zellen damit durch homologe Rekombination

    公开(公告)号：EP0750044A2

    公开(公告)日：1996-12-27

    申请号：EP96202037.6

    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： C12N15/85 ,  A61K48/00 ,  C07K14/505 ,  C07K14/605 ,  C07K14/61

    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 and methods of producing clonal cell strains or heterogenous cell strains.
    The exogenous DNA construct comprises a targeting sequence, a regulatory sequence, an exon and a splice donor site, an intron or a splice acceptor site. A barrier device which permits the passage of the therapeutic product containing the cells and use of the cells in therapeutic applications are also included in the invention.
    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，或本身是所需的（例如治疗性）产品， 转染原代细胞和二次细胞以包括外源性遗传物质，包括通过同源重组进行DNA靶向以及产生克隆细胞株或异源细胞株的方法。 外源DNA构建体包含靶向序列，调控序列，外显子和剪接供体位点，内含子或剪接受体位点。 在治疗应用中允许包含细胞的治疗产物和细胞的使用的阻挡装置也包括在本发明中。 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）。