Comparison of the human versus murine cytomegalovirus immediate early gene promoters for transgene expression by adenoviral vectors
Comparison of the human versus murine cytomegalovirus immediate early gene promoters for transgene expression by adenoviral vectors
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DOI：
10.1099/0022-1317-78-7-1653
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发表时间：
1997-07-01
影响因子：
3.8
通讯作者：
Graham, FL
中科院分区：
文献类型：
--
作者：
Addison, CL;Hitt, M;Graham, FL
我们已经开发了许多复制缺陷型腺病毒（Ad）载体，其在人巨细胞病毒（HCMV）立即早期（IE）基因启动子的控制下表达转基因。在载体骨架中替换E1的表达盒的取向具有显著的转基因表达，定向于Ad基因组右端的表达盒表达比具有相反取向的插入物的表达盒高7倍水平的β-半乳糖苷酶（β-gal）。用其中转基因表达在HCMV IE启动子控制下的几种不同Ad载体中的任何一种感染的鼠细胞产生的转基因产物（如β-gal）比类似感染的人细胞少10-100倍。用鼠CMV（MCMV）IE启动子替换HCMV IE启动子导致在鼠和大鼠细胞中产生的β-gal水平与用HCMV IE启动子获得的水平相比增加约5-30倍，并且与HCMV IE启动子相比，使用MCMV IE启动子在人细胞中产生的水平相同或更高。使用荧光素酶报告基因获得了类似的结果。因此，MCMV IE启动子能够驱动高水平的表达，而没有观察到HCMV IE启动子的明显物种偏好。MCMV IE启动子还在体内指导高水平的表达，这表明携带MCMV IE启动子的Ad载体在动物模型中的转基因表达可能比具有HCMV IE启动子的那些载体更有效。
We have developed a number of replication defective adenoviral (Ad) vectors which express transgenes under the control of the human cytomegalovirus (HCMV) immediate early (IE) gene promoter. The orientation of the expression cassette replacing E1 in the vector backbone had a significant of transgene expression, expression cassettes directed towards the right end of the Ad genome expressing 7-fold higher levels of beta-galactosidase (beta-gal) than those with inserts in the opposite orientation. Murine cells infected with any of several different Ad vectors in which transgene expression was under the control of the HCMV IE promoter produced 10-100-fold less transgene product (such as beta-gal) than similarly infected human cells. Replacing the HCMV IE promoter with the murine CMV (MCMV) IE promoter resulted in an increase in the levels of beta-gal produced in murine and rat cells by approximately 5-30-fold compared to levels obtained with the HCMV IE promoter, and levels produced in human cells were the same or greater using the MCMV IE promoter compared to the HCMV IE promoter. Similar results were obtained using a luciferase reporter gene. The MCMV IE promoter, therefore, was able to drive high levels of expression without the pronounced species preferences observed for the HCMV IE promoter. The MCMV IE promoter also directed high levels of expression in vivo, suggesting that Ad vectors carrying the MCMV IE promoter may be more effective than those with the HCMV IE promoter for transgene expression in animal models.