title: Inhibition of hepatitis B virus gene expression and replication by ribonuclease P.
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.
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, PR China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Xia
authors.forename: Yuan-Chuan
authors.initials: YC
authors.lastname: Chen
authors.forename: Hao
authors.initials: H
authors.lastname: Gong
authors.forename: Wenbo
authors.initials: W
authors.lastname: Zeng
authors.forename: Gia-Phong
authors.initials: GP
authors.lastname: Vu
authors.forename: Phong
authors.initials: P
authors.lastname: Trang
authors.forename: Sangwei
authors.initials: S
authors.lastname: Lu
authors.forename: Jianguo
authors.initials: J
authors.lastname: Wu
authors.forename: Fenyong
authors.initials: F
authors.lastname: Liu
mesh_terms: D000818:Animals; D002460:Cell Line; D015870:Gene Expression; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D016679:Genome, Viral; D006515:Hepatitis B virus / Q000502:physiology*; D006801:Humans; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism / Q000473:pathology; D051379:Mice; D009690:Nucleic Acid Conformation; D058727:RNA, Small Untranslated / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism; D043262:Ribonuclease P / Q000378:metabolism*; D012475:Salmonella / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014779:Virus Replication
chemical_list: D058727:RNA, Small Untranslated; D012367:RNA, Viral; D043262:Ribonuclease P