Evaluation of promoter strength in mouse and rat primary hepatocytes using adenovirus vectors
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- 作者：Eri Arita ; Masuo Kondoh ; Katsuhiro Isoda ; Hikaru Nishimori ; Takeshi Yoshida ; Hiroyuki Mizuguchi ; Kiyohito Yagi
- 关键词：Hepatocyte ; Adenovirus vector ; Promoter ; Transgene activity
- 刊名：European Journal of Pharmaceutics and Biopharmaceutics
- 出版年：2008
- 出版时间：September 2008
- 年：2008
- 卷：70
- 期：1
- 页码：1-6
- 全文大小：374 K
文摘
Primary cultured hepatocytes are widely used in the studies of basic and clinical hepatology. Finding an efficient method for gene transfer into primary hepatocytes will be an important advance for these studies. In the present study, we evaluated the activity of an adenovirus vector including promoters for the Rous sarcoma virus (RSV), elongation factor 1α, and cytomegalovirus (CMV) as well as the β-actin promoter/CMV enhancer (CA) using β-galactosidase as a reporter gene. Although RSV and elongation factor 1α promoters had low transcriptional activity in hepatocytes, the CA and CMV promoters had high activity. The CA promoter was the most active, mediating 50.3- and 204.4-fold more activity than the RSV promoter in mouse and rat hepatocytes, respectively. Dose-response studies revealed that transgene activity can be controlled by as much as 1000-fold, by selection of the promoter and the number of infectious particles per cell. These findings should help in the construction of adenovirus vectors for expressing genes of interest in rodent primary cultured hepatocytes.