抑制乙型肝炎病毒基因的表达和复制的核糖核酸酶P

StephenW

Molecular Therapy (2013); 21 5, 995–1003. doi:10.1038/mt.2013.37
Inhibition of Hepatitis B Virus Gene Expression and Replication by Ribonuclease P

Chuan Xia1, Yuan-Chuan Chen2, Hao Gong3, Wenbo Zeng1, Gia-Phong Vu2, Phong Trang3, Sangwei Lu2,3, Jianguo Wu1 and Fenyong Liu1,2,3

    1State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, P.R. China
    2Program in Comparative Biochemistry, University of California, Berkeley, California, USA
    3School of Public Health, University of California, Berkeley, California, USA

Correspondence: Fenyong Liu, Fenyong Liu, Program in Comparative Biochemistry, University of California, Berkeley, California 94720, USA. E-mail: [email protected]; Jianguo Wu, Jianguo Wu, State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, P.R. China. E-mail: [email protected]; Sangwei Lu, Sangwei Lu, School of Public Health, University of California, Berkeley, California 94720, USA. E-mail: [email protected]

The first three authors contribute equally to this study.

Received 7 November 2012; Accepted 1 February 2013
Advance online publication 12 March 2013
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Abstract

Nucleic acid-based gene interfering approaches, such as those mediated by RNA interference and RNase P-associated external guide sequence (EGS), have emerged as promising antiviral strategies. The RNase P-based technology is unique, because a custom-designed EGS can bind to any complementary mRNA sequence and recruit intracellular RNase P for specific degradation of the target mRNA. In this study, a functional EGS was constructed to target hepatitis B virus (HBV) essential transcripts. Furthermore, an attenuated Salmonella strain was constructed and used for delivery of anti-HBV EGS in cells and in mice. Substantial reduction in the levels of HBV gene expression and viral DNA was detected in cells treated with the Salmonella vector carrying the functional EGS construct. Furthermore, oral inoculation of Salmonella carrying the EGS construct led to an inhibition of ~95% in the levels of HBV gene expression and a reduction of ~200,000-fold in viral DNA level in the livers and sera of the treated mice transfected with a HBV plasmid. Our results suggest that EGSs are effective in inhibiting HBV replication in cultured cells and mammalian livers, and demonstrate the use of Salmonella-mediated delivery of EGS as a promising therapeutic approach for human diseases including HBV infection.

StephenW

基于核酸的基因干扰的方法，如介导的RNA干扰核糖核酸酶P关联的外部引导序列（EGS），已成为有前途的抗病毒策略。基于核糖核酸酶P-技术是独一无二的，因为定制设计的EGS可以绑定到任何互补的mRNA序列特异性降解靶mRNA和招募细胞内的核糖核酸酶P。在这项研究中，一个功能的EGS构建了针对B型肝炎病毒（HBV）必不可少的转录。此外，减毒沙门氏菌菌株，构建了用于在细胞和小鼠抗HBV EGS交付。检测乙型肝炎病毒基因的表达和病毒DNA的水平大幅减少在用沙门氏菌载体的功能的EGS构造的细胞。此外，沙门菌口服接种附带EGS〜95％的抑制HBV基因的表达水平，在接受治疗的小鼠的肝脏和血清中病毒DNA水平与HBV转染〜200000倍的减少导致的构造质粒中。我们的结果表明，的EGSS有效抑制HBV的复制在培养细胞和哺乳动物肝脏，并演示了如何使用EGS是一个很有前途的治疗方法乙肝病毒感染的人类疾病包括沙门氏菌介导的。

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