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Host-vector system for use in gene therapy

机译:用于基因治疗的宿主载体系统

摘要

Host-vector system for expression of a transgene in a target cell or human/animal tissue comprises a eukaryotic cell line transfected with:(a) a recombinant psuedo-retroviral sequence in which all or part of the env gene has been deleted, either totally or partially, and replaced by the transgene as shown in figure 1a, and(b) a nucleic acid sequence which includes a sequence which codes for an envelope protein, is under the control of a promoter, is opt. associated with the transgene and is flanked on its 3' end by a polyadenylation site as shown in figure 1b; provided that(c) both (a) and (b) are capable of trans-complementation to permit the host cell to produce infectious env-deficient virus particles. Also claimed are:(1) a process for the expression of a transgene for gene therapy which comprises:(i) constructing a host-vector system as above;(ii) contacting the system with the cells in which the transgene is to be expressed, and(iii) opt. retransferring the env-contg. sequence into the target cells;(2) a process for the expression of a transgene for gene therapy which comprises the double transfection of target cells with:(i) env-deficient virus particles obtd. using the above host-vector system, and(ii) a nucleic acid sequence contg. a retroviral env gene; where(iii) trans co-expression of the 2 sequences in vivo results in infectious env-deficient recombinant virus particles;(3) a recombinant viral vector contg. a transgene useful in gene therapy, where:(i)a gene essential to the constitution of infectious virus particles is replaced by the transgene;(ii)the essential gene is present on the same or a different vector but is not under the control of viral promoters, and(iii) the prod. of the gene is trans-acting and permits reconstitution of defective virus particles;(4) a medication for use in gene therapy, contg. "vehicles" contg.:(i) a recombinant retroviral sequence in which the env gene is replaced by a trans gene, and(ii) a sequence contg. an env gene under the control of a promoter, where the env-contg. sequence is outside the zone of regulation of the expression of the retroviral sequences, and(5) a medication for use in gene therapy, contg. eukaryotic cells which have been subjected to double transfection and are "represented" by the above host-vector system.
机译:用于在靶细胞或人/动物组织中表达转基因的宿主-载体系统包含真核细胞系,该真核细胞系转染有:(a)重组伪造-逆转录病毒序列,其中全部或部分env基因被完全或全部缺失如图1a所示,部分或部分被转基因替代,并且(b)选择一种核酸序列,该序列包括一个编码包膜蛋白的序列,该序列在启动子的控制下。与转基因结合,并在其3'端侧接一个聚腺苷酸化位点,如图1b所示;前提是(c)(a)和(b)都能够进行反式互补以使宿主细胞产生感染性环境缺陷型病毒颗粒。还要求保护:(1)表达用于基因治疗的转基因的方法,其包括:(i)构建如上的宿主-载体系统;(ii)使该系统与将在其中表达转基因的细胞接触。和(iii)选择。重新传输环境。 (2)表达用于基因治疗的转基因的方法,其包括用以下方法对靶细胞进行双重转染:(i)env缺陷型病毒颗粒obtd。使用上述宿主-载体系统,和(ii)核酸序列续。逆转录病毒env基因; (iii)在体内反式共表达这两个序列会导致感染性环境缺陷的重组病毒颗粒;(3)重组病毒载体。一种可用于基因治疗的转基因,其中:(i)传染性病毒颗粒构成必不可少的基因被该转基因替代;(ii)该必需基因存在于相同或不同的载体上,但不受载体控制病毒启动子,和(iii)产品。该基因的反式作用是反式的,并允许重组有缺陷的病毒颗粒;(4)用于基因治疗的药物,续。 “载体”续:(i)重组en逆转录病毒序列,其中env基因被反式基因替代,和(ii)续序列。在启动子控制下的env基因,其中env-contg。序列位于逆转录病毒序列表达调控区域之外;(5)继续用于基因治疗的药物。经过双重转染并由上述宿主-载体系统“代表”的真核细胞。

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