Characterization of transient expression system for retroviral vector production

被引:19
作者
Hotta, Akitsu
Saito, Yoshikazu
Kyogoku, Kenji
Kawabe, Yoshinori
Nishijima, Ken-ichi
Kamihira, Masamichi
Iijima, Shinji
机构
[1] Nagoya Univ, Dept Biotechnol, Grad Sch Engn, Chikusa Ku, Nagoya, Aichi 4648603, Japan
[2] Kaneka Corp, Takasago, Hyogo 6768688, Japan
基金
日本学术振兴会;
关键词
retroviral vector; murine stem cell virus (MSCV); Q-vector; transient transfection; gene therapy;
D O I
10.1263/jbb.101.361
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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.
引用
收藏
页码:361 / 368
页数:8
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