title: Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro.
abstract: OBJECTIVE
  To explore the anti-hepatitis B virus effect of RNA interference (RNAi) using small hairpin RNA (shRNA) expression vector.
  
  
  METHODS
  Hepatitis B virus surface antigen green fluorescent protein (HBs-GFP) fusion vector and shRNA expression vectors were constructed and cotransfected transiently into HepG2 cells. mRNAs extracted from HepG2 cells were detected by real-time PCR. Fluorescence of HBs-GFP protein was detected by fluorescence-activated cell sorting (FACS). The effective shRNA expression vector was transfected into HepG2.2.15 cells. HBsAg and HBeAg in HepG2.2.15 cells were analyzed by radioimmunoassay (RIA) method.
  
  
  RESULTS
  FACS revealed that shRNA targeting at HBsAg reduced the GFP signal by 56% compared to the control. Real-time PCR showed that HBs-GFP mRNA extracted from HepG2 cells cotransfected with pAVU6+27 and HBs-GFP expression plasmids decreased by 90% compared to the empty vector control. The expressions of HBsAg and HBeAg were also inhibited by 43% and 64%, respectively.
  
  
  CONCLUSIONS
  RNAi using shRNA expression vector can inhibit the expression of HBsAg, providing a fresh approach to screening the efficient small interfering RNAs (siRNAs).
authors.affiliation: Institute of Infectious Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, Zhejiang Province, China.
authors.forename: Zheng-Gang
authors.initials: ZG
authors.lastname: Yang
authors.forename: Zhi
authors.initials: Z
authors.lastname: Chen
authors.forename: Qin
authors.initials: Q
authors.lastname: Ni
authors.forename: Ning
authors.initials: N
authors.lastname: Xu
authors.forename: Jun-Bin
authors.initials: JB
authors.lastname: Shao
authors.forename: Hang-Ping
authors.initials: HP
authors.lastname: Yao
mesh_terms: D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D006513:Hepatitis B e Antigens / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D008113:Liver Neoplasms; D010957:Plasmids; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
chemical_list: D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins