Barry S C, Harder B, Brzezinski M, Flint L Y, Seppen J, Osborne W R
Department of Pediatrics, University of Washington, Seattle, WA 98195, USA.
Hum Gene Ther. 2001 Jun 10;12(9):1103-8. doi: 10.1089/104303401750214311.
Incorporation of a central polypurine tract (cPPT) and a posttranscriptional regulatory element (PRE) into lentivirus vectors provides increased transduction efficiency and transgene expression. We compared the effects of these elements individually and together on transduction efficiency and gene expression, using lentivirus vectors pseudotyped with vesicular stomatitis virus G protein (VSV-G) and encoding enhanced green fluorescent protein (GFP) and rat erythropoietin (EPO). The transduction efficiency was greater than 2-fold higher in the vector containing the PRE element, 3-fold higher in vector encoding the cPPT element, and 5-fold increased in the GFP virus containing both cPPT and PRE elements relative to the parent virus. In comparison with parent vector the mean fluorescence intensity (MFI) of GFP expression was 7-fold higher in cells transduced with virus containing PRE, 6-fold increased in cells transduced with virus containing cPPT, and 42-fold increased in GFP-virus containing both cPPT and PRE elements. EPO-virus containing a PRE element showed a nearly 5-fold increase in EPO secretion over the parent vector, and the vector encoding both PRE and cPPT showed a 65-fold increase. Thus, lentivirus vectors incorporating both PRE and cPPT showed expression levels significantly increased over the sum of the components alone, suggesting a synergistic effect.
将中央多嘌呤序列（cPPT）和转录后调控元件（PRE）整合到慢病毒载体中可提高转导效率和转基因表达。我们使用伪型为水泡性口炎病毒G蛋白（VSV-G）并编码增强型绿色荧光蛋白（GFP）和大鼠促红细胞生成素（EPO）的慢病毒载体，比较了这些元件单独及共同作用对转导效率和基因表达的影响。相对于亲本病毒，含有PRE元件的载体转导效率提高了2倍以上，编码cPPT元件的载体转导效率提高了3倍，同时含有cPPT和PRE元件的GFP病毒转导效率提高了5倍。与亲本载体相比，用含PRE的病毒转导的细胞中GFP表达的平均荧光强度（MFI）提高了7倍，用含cPPT的病毒转导的细胞中提高了6倍，同时含有cPPT和PRE元件 的GFP病毒转导的细胞中提高了42倍。含有PRE元件的EPO病毒的EPO分泌比亲本载体增加了近5倍，同时编码PRE和cPPT的载体增加了65倍。因此，同时含有PRE和cPPT的慢病毒载体的表达水平比单独各元件之和显著提高，表明存在协同效应。
