Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA
Inhibition of hepatitis C virus infection and expression in vitro and in vivo by recombinant adenovirus expressing short hairpin RNA
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DOI：
10.1111/j.1440-1746.2007.05076.x
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发表时间：
2008-09-01
影响因子：
4.1
通讯作者：
Watanabe, Mamoru
中科院分区：
文献类型：
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作者：
Sakamoto, Naoya;Tanabe, Yoko;Watanabe, Mamoru
背景和目的：我们以前曾报道，合成的小干扰RNA（siRNA）和基于DNA的siRNA表达载体有效地和特异性地抑制丙型肝炎病毒（HCV）的复制在体外。方法：构建表达短发夹状RNA（short hairpin RNA，shRNA）的重组逆转录病毒和腺病毒，体外转染复制子表达细胞，体内转染转基因小鼠，观察siRNA对HCV RNA的抑制作用。逆转录病毒转导Huh 7细胞以表达shRNA，随后将HCV复制子转染到细胞中，表明细胞已获得对HCV的抗性复制的用表达shRNA的腺病毒感染表达HCV复制子的细胞导致有效的载体递送和shRNA的表达，导致细胞中复制子的抑制类似于10（-3）。将表达shRNA的腺病毒静脉注射到可通过Cre/loxP转换系统诱导表达HCV结构蛋白的转基因小鼠中，导致肝脏中病毒蛋白合成的特异性抑制。总之，我们的结果支持利用基于siRNA和/或shRNA表达的基因靶向治疗来对抗HCV复制的可行性，可能对治疗丙型肝炎有价值
Background and Aim: We have reported previously that synthetic small interfering RNA (siRNA) and DNA-based siRNA expression vectors efficiently and specifically suppress hepatitis C virus (HCV) replication in vitro. In this study, we investigated the effects of the siRNA targeting HCV-RNA in vivo.Methods: We constructed recombinant retrovirus and adenovirus expressing short hairpin RNA (shRNA), and transfected into replicon-expressing cells in vitro and transgenic mice in vivo.Results: Retroviral transduction of Huh7 cells to express shRNA and subsequent transfection of an HCV replicon into the cells showed that the cells had acquired resistance to HCV replication. Infection of cells expressing the HCV replicon with an adenovirus expressing shRNA resulted in efficient vector delivery and expression of shRNA, leading to suppression of the replicon in the cells by similar to 10(-3). Intravenous delivery of the adenovirus expressing shRNA into transgenic mice that can be induced to express HCV structural proteins by the Cre/loxP switching system resulted in specific suppression of virus protein synthesis in the liver.Conclusion: Taken together, our results support the feasibility of utilizing gene targeting therapy based on siRNA and/or shRNA expression to counteract HCV replication, which might prove valuable in the treatment of hepatitis C.