基于逆转录病毒递送的RNA干扰对HepG2.2.15细胞中乙型肝炎病毒表达和复制的稳定抑制

Stable inhibition of hepatitis B virus expression and replication in HepG2.2.15 cells by RNA interference based on retrovirus delivery.

作者信息

Jia Fang, Zhang Yi-Zheng, Liu Chang-Mei

机构信息

Molecular Virology Research Center, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.

出版信息

J Biotechnol. 2007 Jan 30;128(1):32-40. doi: 10.1016/j.jbiotec.2006.09.007. Epub 2006 Sep 23.

DOI:10.1016/j.jbiotec.2006.09.007

PMID:17049658

Abstract

RNA interference (RNAi) of virus-specific genes has emerged as a potential antiviral strategy. In order to suppress hepatitis B virus (HBV) expression and replication, a retrovirus-based RNAi system was developed, which utilized the U6-RNA polymerase III (Pol III) promoter to drive efficient expression and deliver the HBV-specific short hairpin RNAs (shRNAs) in HepG2.2.15 (2215) cells. In this system, the retrovirus vector with a puromycin selection marker was integrated into the host cell genome and allowed stable expression of shRNAs. In Puro-resistant 2215 cells, the levels of both HBV protein and mRNA were dramatically reduced by over 88% and HBV replication was suppressed. The results demonstrated that retrovirus-based RNAi technology will have foreseeable applications both in experimental biology and molecular medicine.

摘要

病毒特异性基因的RNA干扰（RNAi）已成为一种潜在的抗病毒策略。为了抑制乙型肝炎病毒（HBV）的表达和复制，开发了一种基于逆转录病毒的RNAi系统，该系统利用U6-RNA聚合酶III（Pol III）启动子驱动高效表达，并在HepG2.2.15（2215）细胞中递送HBV特异性短发夹RNA（shRNA）。在该系统中，带有嘌呤霉素选择标记的逆转录病毒载体整合到宿主细胞基因组中，并允许shRNA稳定表达。在对嘌呤霉素耐药的2215细胞中，HBV蛋白和mRNA水平均显著降低超过88%，HBV复制受到抑制。结果表明，基于逆转录病毒的RNAi技术在实验生物学和分子医学中都将有可预见的应用。