Stable inhibition of hepatitis B virus expression and replication in HepG2.2.15 cells by RNA interference based on retrovirus delivery.
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.
Jia
Fang
F
Molecular Virology Research Center, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.
Zhang
Yi-Zheng
YZ
Liu
Chang-Mei
CM
D015967:Gene Expression Regulation, Viral / Q000235:genetics*; D005822:Genetic Vectors; D006509:Hepatitis B / Q000517:prevention & control*; D006515:Hepatitis B virus / Q000235:genetics / Q000502:physiology*; D006801:Humans; D034622:RNA Interference; D012190:Retroviridae / Q000235:genetics* / Q000472:pathogenicity; D014407:Tumor Cells, Cultured; D014779:Virus Replication