Characterization of transient expression system for retroviral vector production
Characterization of transient expression system for retroviral vector production
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DOI：
10.1263/jbb.101.361
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发表时间：
2006-04-01
影响因子：
2.8
通讯作者：
Iijima, Shinji
中科院分区：
文献类型：
--
作者：
Hotta, Akitsu;Saito, Yoshikazu;Iijima, Shinji
已经改进了使用瞬时表达系统生产逆转录病毒载体以获得高滴度病毒制剂，由于转基因的细胞毒性或细胞抑制作用，使用包装细胞系难以生产该高滴度病毒制剂。在这里，我们使用了一种这样的生产方法，即所谓的Q-载体系统，并研究了其生产病毒的潜力。与包装细胞系统相比，Q载体系统可以产生类似水平的病毒载体，但产量似乎取决于转基因的大小和性质。在研究瞬时表达系统和包装细胞系统之间病毒组分的定量差异的过程中，我们发现与包装细胞系统相比，Q-载体系统可以表达更高量的病毒RNA和蛋白。然而，与包装细胞系统产生的病毒滴度相比，这并没有导致更高的病毒滴度。这表明在瞬时系统中从质粒转录的逆转录病毒RNA似乎主要用于翻译，并且只有一些RNA分子包装在病毒颗粒中。
The production of retroviral vectors using a transient expression system has been improved to obtain a high-titer virus preparation that is difficult to produce using packaging cell lines due to the cytotoxic or cytostatic effect of transgenes. Here, we used one such production method, the so-called Q-vector system, and examined its potential for virus production. The Q-vector system could produce a similar level of viral vectors compared with the packaging cell system but the production seemed to depend on the size and nature of transgenes. In the process of investigation of the quantitative difference in viral components between the transient expression system and the packaging cell system, we found that the Q-vector system could express higher amounts of viral RNA and proteins compared with the packaging cell system. However, this did not lead to a higher virus titer compared with that produced by the packaging cell system. This suggests that retroviral RNA transcribed from the plasmid in the transient system seemed to be used mainly for translation and only some of the RNA molecules were packaged in viral particles.