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    GENETIC MODIFICATION OF RATS
    5.
    发明申请
    GENETIC MODIFICATION OF RATS 审中-公开
    RATS的遗传修饰

    公开(公告)号：US20170037429A1

    公开(公告)日：2017-02-09

    申请号：US15242025

    申请日：2016-08-19

    申请人： Regeneron Pharmaceuticals, Inc.

    发明人： Jeffrey D. Lee ,  Wojtek Auerbach ,  David Heslin ,  David Frendewey ,  Ka-Man Venus Lai ,  David M. Valenzuela

    IPC分类号： C12N15/85 ,  A01K67/02 ,  C12N15/877 ,  A01K67/027

    CPC分类号： C12N15/8509 ,  A01K67/02 ,  A01K67/0271 ,  A01K67/0275 ,  A01K2207/12 ,  A01K2217/15 ,  A01K2227/105 ,  A61D19/04 ,  C12N5/0606 ,  C12N15/8775 ,  C12N2501/235 ,  C12N2501/727 ,  C12N2501/999 ,  C12N2510/00

    摘要： Compositions and methods are provided for making rat pluripotent and totipotent cells, including rat embryonic stem (ES) cells. Compositions and methods for improving efficiency or frequency of germline transmission of genetic modifications in rats are provided. Such methods and compositions comprise an in vitro culture comprising a feeder cell layer and a population of rat ES cells or a rat ES cell line, wherein the in vitro culture conditions maintain pluripotency of the ES cell and comprises a media having mouse leukemia inhibitory factor (LIF) or an active variant or fragment thereof. Various methods of establishing such rat ES cell lines are further provided. Methods of selecting genetically modified rat ES cells are also provided, along with various methods to generate a transgenic rat from the genetically modified rat ES cells provided herein. Various kits and articles of manufacture are further provided.

    摘要翻译： 提供组合物和方法用于制备大鼠多能全细胞，包括大鼠胚胎干（ES）细胞。 提供了用于提高大鼠遗传修饰的种系传播效率或频率的组合物和方法。 这样的方法和组合物包括体外培养物，其包含饲养细胞层和大鼠ES细胞群或大鼠ES细胞系，其中体外培养条件保持ES细胞的多能性，并且包含具有小鼠白血病抑制因子 LIF）或其活性变体或片段。 还提供了建立这种大鼠ES细胞系的各种方法。 还提供了选择遗传修饰的大鼠ES细胞的方法，以及从本文提供的遗传修饰的大鼠ES细胞产生转基因大鼠的各种方法。 还提供了各种套件和制品。

    Methods for mitochondrial DNA replacement in oocytes
    6.
    发明授权
    Methods for mitochondrial DNA replacement in oocytes 有权
    卵母细胞线粒体DNA置换方法

    公开(公告)号：US09434921B2

    公开(公告)日：2016-09-06

    申请号：US13265326

    申请日：2010-04-22

    申请人： Shoukhrat M. Mitalipov

    发明人： Shoukhrat M. Mitalipov

    IPC分类号： A01K67/027 ,  C12N15/00 ,  C12N5/075 ,  C12N5/16 ,  C12N15/877 ,  C12N15/873 ,  C12N5/073

    CPC分类号： C12N5/0609 ,  C12N5/0604 ,  C12N5/16 ,  C12N15/873 ,  C12N15/8775 ,  C12N15/8776 ,  C12N2517/10

    摘要： Methods are provided for producing a primate oocyte in vitro. The methods include removing nuclear DNA from a recipient primate oocyte from a first primate in a manner that does not lower levels of maturation promoting factor (MPF) to form an enucleated recipient primate oocyte. The recipient primate oocyte is enucleated using a non-UV-based spindle imaging system. Nuclear genetic material or DNA including chromosomes from a donor primate oocyte arrested at metaphase II from a second primate is isolated in the form of the karyoplast and introduced into the enucleated recipient primate oocyte. Introduction of the chromosomes is performed using a fusogenic agent or electroporation to produce a hybrid oocyte.

    摘要翻译： 提供了用于在体外产生灵长类动物卵母细胞的方法。 所述方法包括以不降低成熟促进因子（MPF）水平以形成去核的受体灵长类动物卵母细胞的方式从第一灵长类动物中移除来自受体灵长类动物卵母细胞的核DNA。 使用非基于UV的主轴成像系统使受体灵长类动物卵母细胞被去核。 来自第二种灵长类动物的中期II停止的供体灵长类卵母细胞的核遗传物质或DNA包括染色体，以核型质粒的形式分离，并引入去核的受体灵长类动物卵母细胞。 使用融合剂或电穿孔来进行染色体的引入以产生杂交卵母细胞。