A retroviral vector was constructed with an autoregulatory cassette to allow expression of the gene of interest in response to oral administration of doxycycline (Dox) in vivo. The cassette contains all the components of the reverse tetracycline-regulated (rtTA) system, a drug selectable marker with the internal ribosome entry site and the gene of interest (GFP). FACS analyses showed an induction of GFP-fluorescence of two orders of magnitude in retrovirus-infected 208F cells dependent on the amount of Dox in the medium. Furthermore, oral administration of Dox resulted in GFP expression in transplanted 208F cells in the peritoneal cavity of nude mice. Thus this reverse tetracycline-regulated retroviral vector (RTRV) system simplifies the delivery of controllable genes to cultured and implanted cells. It is hoped that this approach may pave the way to controlled gene expression during a particular window of time in gene therapy applications.

译文

用自动调节盒构建逆转录病毒载体,以响应体内口服强力霉素 (Dox) 的目的基因表达。该盒包含反向四环素调节 (rtTA) 系统的所有成分,该系统是具有内部核糖体进入位点和目标基因 (GFP) 的药物选择标记。FACS分析显示,在逆转录病毒感染的208F细胞中,取决于培养基中Dox的量,诱导了两个数量级的GFP荧光。此外,口服Dox导致裸鼠腹膜腔中移植的208F细胞中GFP表达。因此,这种反向四环素调节的逆转录病毒载体 (RTRV) 系统简化了可控基因向培养和植入细胞的传递。希望这种方法可以为在基因治疗应用中的特定时间窗口内控制基因表达铺平道路。

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