A GENERAL-METHOD FOR THE GENERATION OF HIGH-TITER, PANTROPIC RETROVIRAL VECTORS - HIGHLY EFFICIENT INFECTION OF PRIMARY HEPATOCYTES
A GENERAL-METHOD FOR THE GENERATION OF HIGH-TITER, PANTROPIC RETROVIRAL VECTORS - HIGHLY EFFICIENT INFECTION OF PRIMARY HEPATOCYTES
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DOI：
10.1073/pnas.91.20.9564
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发表时间：
1994-09-27
影响因子：
11.1
通讯作者：
FRIEDMANN, T
中科院分区：
文献类型：
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作者：
YEE, JK;MIYANOHARA, A;FRIEDMANN, T
逆转录病毒载体一直是导致人类基因治疗的许多研究的核心组成部分。然而，逆转录病毒载体在人类细胞中的普遍低滴度和低效感染性限制了它们的使用。我们以前报道过水疱性口炎病毒的G蛋白可以作为表达水疱性口炎病毒G的特定逆转录病毒载体LGRNL的唯一包膜蛋白成分。现在我们报道了一种更有用的通用瞬时转染方案，可以从任何基于Moloney小鼠白血病的逆转录病毒载体中产生非常高滴度的水泡性口炎病毒G包膜伪型，而不必依赖逆转录病毒载体本身的细胞毒性G蛋白的表达。我们还证明了假型LacZ载体在原代小鼠肝细胞中的感染效率非常高。我们建议，携带报告基因的伪型逆转录病毒载体将允许在许多以前无法接触到的脊椎动物和无脊椎动物系统中进行基因研究。此外，由于这些载体代表了足够高滴度的逆转录病毒载体，从而允许有效的直接逆转录病毒介导的体内基因转移，我们还建议，携带潜在治疗性基因的伪型载体将成为测试体内基因治疗潜力的有用工具。
Retroviral vectors have been central components in many studies leading to human gene therapy. However, the generally low titers and inefficient infectivity of retroviral vectors in human cells have limited their use. We previously reported that the G protein of vesicular stomatitis virus can serve as the exclusive envelope protein component for one specific retroviral vector, LGRNL, that expresses vesicular stomatitis virus G. We now report a more useful general transient transfection scheme for producing very high-titer vesicular stomatitis virus G-enveloped pseudotypes from any Moloney murine leukemia-based retroviral vector without having to rely on the expression of the cytotoxic G protein from the retroviral vector itself. We also demonstrate very high efficiency of infection with a pseudotyped lacZ vector in primary mouse hepatocytes. We suggest that pseudotyped retroviral vectors carrying reporter genes will permit genetic studies in many previously inaccessible vertebrate and invertebrate systems. Furthermore, because these vectors represent retroviral vectors of sufficiently high titer to allow efficient direct retroviral-mediated in vivo gene transfer, we also suggest that pseudotyped vectors carrying potentially therapeutic genes will become useful to test the potential for in vivo gene therapy.