Burns J C, Friedmann T, Driever W, Burrascano M, Yee J K
Department of Pediatrics, University of California, San Diego School of Medicine, La Jolla 92093.
Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8033-7. doi: 10.1073/pnas.90.17.8033.
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.
逆转录病毒载体的宿主细胞范围受限以及滴度较低,这限制了它们在真核细胞中进行稳定基因转移的应用。为克服这些限制,我们构建了源自鼠白血病病毒的载体,其中逆转录病毒包膜糖蛋白已完全被水泡性口炎病毒的G糖蛋白所取代。此类载体可通过超速离心浓缩至滴度>10(9)集落形成单位/毫升,并且能够感染通常对含有逆转录病毒包膜蛋白的载体感染具有抗性的细胞,如仓鼠和鱼类细胞系。浓缩水泡性口炎病毒G糖蛋白假型载体的能力将有助于基因治疗模型研究以及其他需要在体内直接递送载体的基因转移实验。这些假型载体的可得性还将通过高效导入和表达外源基因,促进非哺乳动物物种的遗传学研究,包括重要的斑马鱼发育系统。