PRODUCTION OF HIGH-TITER HELPER-FREE RETROVIRUSES BY TRANSIENT TRANSFECTION
PRODUCTION OF HIGH-TITER HELPER-FREE RETROVIRUSES BY TRANSIENT TRANSFECTION
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DOI：
10.1073/pnas.90.18.8392
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发表时间：
1993-09-15
影响因子：
11.1
通讯作者：
BALTIMORE, D
中科院分区：
文献类型：
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作者：
PEAR, WS;NOLAN, GP;BALTIMORE, D
通过使用高转染效率的293T细胞系，并向其中稳定导入表达逆转录病毒包装功能的构建体，利用瞬时转染实现了高滴度、无辅助病毒的逆转录病毒的产生。由此产生的嗜亲性病毒包装细胞系BOSC 23在磷酸钙介导的转染后72小时内，可在上清液中产生>10⁶感染单位/毫升的感染性逆转录病毒。对具有复制能力的病毒进行的严格检测表明不存在辅助病毒。该系统能够产生高滴度的逆转录病毒载体，这些载体所表达的基因在稳定的生产细胞系中极难以高滴度繁殖。这种方法应促进和扩大无辅助病毒的逆转录病毒基因转移的应用，并且对基因治疗有用。
The generation of high-titer, helper-free retroviruses by transient transfection has been achieved by using the highly transfectable 293T cell line into which are stably introduced constructs that express retroviral packaging functions. The resulting ecotropic virus packaging cell line BOSC 23 produces infectious retrovirus at >10(6) infectious units/ml of supernatant within 72 hr after CaPO4-mediated transfection. A stringent assay for replication-competent virus showed that no helper virus was present. The system can produce high titers of retroviral vectors expressing genes that are extremely difficult to propagate at high titer in stable producer lines. This method should facilitate and extend the use of helper-free retroviral gene transfer, as well as be useful for gene therapy.