Episomal vectors rapidly and stably produce high-titer recombinant retrovirus
Episomal vectors rapidly and stably produce high-titer recombinant retrovirus
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DOI：
10.1089/hum.1996.7.12-1405
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发表时间：
1996-08-01
影响因子：
4.2
通讯作者：
Nolan, GP
中科院分区：
文献类型：
--
作者：
Kinsella, TM;Nolan, GP
EB病毒（EBV）的核复制和保留功能已在此用于在基于人细胞的逆转录病毒包装系内维持附加型逆转录病毒构建体。这些杂合EBV/逆转录病毒构建体能够在转染到基于293 T的亲嗜性和/或亲嗜性逆转录病毒包装细胞中后48小时内产生无辅助基因的重组逆转录病毒，并持续至少30天。在嘌呤霉素选择转染的逆转录病毒包装细胞后，仅获得大于10（7）TU/ml的病毒滴度，这种附加型逆转录病毒生产方法避免了瞬时和染色体稳定的逆转录病毒生产系统中固有的一些限制，提供了可重复的快速、大规模、稳定和高滴度逆转录病毒生产。
The nuclear replication and retention functions of the Epstein-Barr virus (EBV) have been utilized here to maintain retroviral constructs episomally within human cell-based retroviral packaging lines. These hybrid EBV/retroviral constructs are capable of producing helper-free recombinant retrovirus as soon as 48 hr and for at least 30 days after transfection into 293T-based ecotropic and/or amphotropic retroviral packaging cells, Viral titers greater than 10(7) TU/ml mere obtained after puromycin selection of transfected retroviral packaging cells, This episomal approach to retroviral production circumvents some limitations inherent in transient and chromosomally stable retroviral producer systems, affording reproducibly rapid, large-scale, stable, and high-titer retrovirus production.