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    • 8. 发明申请
    • UTILIZATION OF NON-VIRAL SEQUENCES FOR MINUS-STRAND DNA TRANSFER AND GENE RECONSTITUTION
    • 非转基因DNA转移和基因重组的非病毒序列的利用
    • WO0190391A3
    • 2002-04-11
    • PCT/US0115739
    • 2001-05-15
    • US HEALTHHU WEI SHAUPATHAK VINAY K
    • HU WEI-SHAUPATHAK VINAY K
    • A61K38/00A61K48/00C12N15/867C12N15/90C12N15/86
    • C12N15/86A61K38/00A61K48/00C12N15/907C12N2740/13043C12N2830/50C12N2840/20C12N2840/203
    • A retroviral vector for gene reconstitution is provided that includes a 3' portion of a heterologous nucleic acid sequence 5' of a first att site of the retroviral vector, a 5' portion of the heterologous nucleic acid sequence 3' of a second att site of the retroviral vector. In this vector, a sub-portion of the 3' portion of the heterologous nucleic acid sequence and the 5' region of the heterologous nucleic acid sequence are direct repeats. Transformation of a eukaryotic cell with the retroviral vector results in reconstitution and duplication of the heterologous nucleic acid sequence. A retroviral vector for gene reconsitution is also provided that includes a 3' portion of a heterologous nucleic acid sequence inserted into or adjacent to a 5' retroviral terminal repeat of the retroviral vector, and a 5'portion of the heterologous nucleic acid sequence inserted into or adjacent to a 3' retroviral terminal repeat of the retroviral vector, wherein the 5' retroviral terminal repeat and the 3' retroviral terminal repeat each comprise an att site. A sub-portion of the the 3' portion of the heterologous nucleic acid sequence and the 5' region of the heterologous nucleic acid sequence are direct repeats. Transformation of a eukaryotic cell with this retroviral vector results in reconstitution and duplication of the heterologous nucleic acid sequence. Method and kits are provided for reconstituting and duplicating a nucleic acid molecule in a host cell using these retroviral vectors to introduce a heterologous nucleic acid in the cell. A method is provided for treating a subject that includes contacting a cell of the subject with a therapeutically effective amount of a retroviral vector for gene reconstitution and duplication.
    • 提供了用于基因重构的逆转录病毒载体,其包含逆转录病毒载体的第一个att位点的异源核酸序列5'的3'部分,第二个att位点的异源核酸序列3'的5'部分 逆转录病毒载体。 在该载体中,异源核酸序列的3'部分的亚部分和异源核酸序列的5'区域是直接重复的。 用逆转录病毒载体转化真核细胞导致异源核酸序列的重组和重复。 还提供了用于基因重新取代的逆转录病毒载体,其包括插入或邻近逆转录病毒载体的5'逆转录病毒末端重复序列的异源核酸序列的3'部分和插入到其中的异源核酸序列的5'部分 或与逆转录病毒载体的3'逆转录病毒终端重复相邻,其中5'逆转录病毒终端重复和3'逆转录病毒末端重复各自包含att位点。 异源核酸序列的3'部分的亚部分和异源核酸序列的5'区域是直接重复的。 用该逆转录病毒载体转化真核细胞导致重组和重复异源核酸序列。 提供了使用这些逆转录病毒载体在宿主细胞中重构和复制核酸分子以在细胞中引入异源核酸的方法和试剂盒。 提供了一种治疗受试者的方法,其包括使受试者的细胞与治疗有效量的逆转录病毒载体接触,用于基因重组和复制。