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Vesicular stomatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells.

机译:水泡性口炎病毒G糖蛋白假型逆转录病毒载体:高滴度浓缩并有效地将基因转移到哺乳动物和非哺乳动物细胞中。

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摘要

The restricted host-cell range and low titer of retroviral vectors limit their use for stable gene transfer in eukaryotic cells. To overcome these limitations, we have produced murine leukemia virus-derived vectors in which the retroviral envelope glycoprotein has been completely replaced by the G glycoprotein of vesicular stomatitis virus. Such vectors can be concentrated by ultracentrifugation to titers > 10(9) colony-forming units/ml and can infect cells, such as hamster and fish cell lines, that are ordinarily resistant to infection with vectors containing the retroviral envelope protein. The ability to concentrate vesicular stomatitis virus G glycoprotein pseudotyped vectors will facilitate gene therapy model studies and other gene transfer experiments that require direct delivery of vectors in vivo. The availability of these pseudotyped vectors will also facilitate genetic studies in nonmammalian species, including the important zebrafish developmental system, through the efficient introduction and expression of foreign genes.
机译:受限的宿主细胞范围和低滴度的逆转录病毒载体限制了它们在真核细胞中稳定基因转移的用途。为了克服这些限制,我们生产了鼠白血病病毒衍生的载体,其中逆转录病毒包膜糖蛋白已被水泡性口炎病毒的G糖蛋白完全取代。可以通过超速离心将这些载体浓缩至滴度> 10(9)集落形成单位/ ml,并且可以感染通常对含有逆转录病毒包膜蛋白的载体抗感染的细胞,例如仓鼠和鱼细胞系。浓缩水泡性口腔炎病毒G糖蛋白假型载体的能力将促进基因治疗模型研究和其他需要在体内直接递送载体的基因转移实验。这些假型载体的可用性还将通过有效引入和表达外源基因,促进非哺乳动物物种(包括重要的斑马鱼发育系统)的遗传研究。

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