Beaudoin E L, Bais A J, Junghans R P
Boston University School of Medicine, Department of Surgical Research, North Campus, Roger Williams Medical Center, 825 Chalkstone Avenue, Providence, RI 02908, USA.
J Virol Methods. 2008 Mar;148(1-2):253-9. doi: 10.1016/j.jviromet.2007.12.008. Epub 2008 Feb 4.
Vector producer cells are derived from helper cell lines expressing viral proteins that have been transduced to express a transgene-carrying retroviral genome. Vector producing cells express two relevant forms of RNA in their cytoplasm: vector RNA (vRNA) that is packaged as the actual gene transfer agent, and messenger RNA (mRNA) from which transgene is translated. Two premises underlie this study: (1) vRNA is limiting for virus production and (2) mRNA is proportional to vRNA. Together, these premises predict that transgene expression in the vector producing cells will be predictive of the viral titer from those cells. In this case, sorting the vector producing cells for high transgene expression should select for more virus production in vector producing cell supernatants. This prediction was supported, with a greater than fivefold benefit in viral titer. This demonstrates a rapid and simple method by which to obtain significantly increased viral titers from the same vector producing cell preparation.
载体生产细胞源自表达病毒蛋白的辅助细胞系，这些辅助细胞系已被转导以表达携带转基因的逆转录病毒基因组。载体生产细胞在其细胞质中表达两种相关形式的RNA：作为实际基因传递剂被包装的载体RNA（vRNA），以及用于翻译转基因的信使RNA（mRNA）。本研究基于两个前提：（1）vRNA限制病毒产生，（2）mRNA与vRNA成比例。总之，这些前提预测载体生产细胞中的转基因表达将预示这些细胞的病毒滴度。在这种情况下，对载体生产细胞进行高转基因表达分选应能选择出在载体生产细胞上清液中产生更多病毒的细胞。这一预测得到了支持，病毒滴度提高了五倍以上。这证明了一种快速且简单的方法，可从相同的载体生产细胞制剂中获得显著提高的病毒滴度。
