Evaluation of promoter strength in mouse and rat primary hepatocytes using adenovirus vectors
Evaluation of promoter strength in mouse and rat primary hepatocytes using adenovirus vectors
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DOI：
10.1016/j.ejpb.2008.03.013
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发表时间：
2008-09-01
影响因子：
4.9
通讯作者：
Yagi, Kiyohito
中科院分区：
文献类型：
--
作者：
Arita, Eri;Kondoh, Masuo;Yagi, Kiyohito
原代培养的肝细胞广泛应用于基础和临床肝病学研究。寻找一种将基因转移到原代脂肪细胞的有效方法将是这些研究的重要进展。在本研究中，我们使用β-半乳糖苷酶Lis报告基因评估了腺病毒载体的活性，该载体包括劳斯肉瘤病毒（RSV）、延伸因子1α和巨细胞病毒（CMV）的启动子以及P-肌动蛋白启动子/CMV增强子（CA）。尽管RSV和延伸因子批次启动子在肝细胞中具有低转录活性，但CA和CMV启动子具有高活性。 CA启动子是最活跃的，在小鼠和大鼠肝细胞中介导的活性分别比RSV启动子高50.3倍和204.4倍。剂量反应研究表明，通过选择启动子和每个细胞的感染颗粒数量，转基因活性可控制多达 1000 倍。这些发现应有助于构建腺病毒载体，用于在啮齿动物原代培养的肝细胞中表达感兴趣的基因。 (c) 2008 Elsevier B.V. 保留所有权利。
Primary cultured hepatocytes are widely used in the Studies or basic and clinical hepatology. Finding an efficient method for gene transfer into primary licpatocytes will be ail 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 1 alpha, and cytomegalovirus (CMV) as well Lis the P-actin promoter/CMV enhancer (CA) using beta-galactosidase Lis a reporter gene. Although RSV and elongation factor lot 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. (c) 2008 Elsevier B.V. All rights reserved.