title: Optimized tandem amiRNA mediates stronger inhibitory effects on hepatitis B virus infection.
abstract: OBJECTIVE
  RNA interference (RNAi) has emerged as a potential new approach against hepatitis B virus (HBV) infection but unfortunately it also selects resistant virus mutants. In this study we combined the advantages of artificial micro RNAs (amiRNAs) reported previously with the purpose of constructing a more practical amiRNA with high inhibition effects against HBV.
  
  
  METHODS
  Aiming at conserved sites, we constructed singular-sequence vectors amiRNA-HBV1, amiRNA-HBV2, amiRNA-HBV3 and amiRNA-HBV4. We chose the two sequences of high efficiency, then built the tandem-sequence vector amiRNA-HBV3-HBV4. These vectors were transfected into HepG2.2.15 transiently. The secreted HBV surface antigen (HBsAg) and HBV 'e' antigen (HBeAg) were measured with a chemiluminescent microparticle immunoassay, and intracellular and extracellular HBV DNA was quantified by real-time PCR.
  
  
  RESULTS
  Our results demonstrated that amiRNA-HBV1, amiRNA-HBV2, amiRNA-HBV3, and amiRNA-HBV4 achieved a maximum inhibition of HBV mRNA expression of 29.3%, 14.9%, 61.2%, and 75.6%, respectively, while the tandem amiRNA-HBV3-HBV4 vector led to an inhibition of 87.2%.
  
  
  CONCLUSIONS
  Taken together, our data suggest that vector-based multiple artificial microRNAs are a promising therapeutic approach for chronic HBV infection.
authors.affiliation: Dalian Sixth People's Hospital, Dalian 116033, China.
authors.forename: Chunwen
authors.initials: C
authors.lastname: Pu
authors.forename: Lifen
authors.initials: L
authors.lastname: Wang
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Miao
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chunmeng
authors.initials: C
authors.lastname: Jiang
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Weixiang
authors.initials: W
authors.lastname: Sun
authors.forename: Qingwei
authors.initials: Q
authors.lastname: Gao
mesh_terms: D001483:Base Sequence; D004261:DNA Replication; D004279:DNA, Viral / Q000032:analysis; D005822:Genetic Vectors; D056945:Hep G2 Cells; D006509:Hepatitis B / Q000628:therapy*; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D034622:RNA Interference; D014162:Transfection
chemical_list: D004279:DNA, Viral; D035683:MicroRNAs