[Potent and specific inhibition of hepatitis B virus antigen expression by RNA interference].
OBJECTIVE
To develop an RNAi approach that specifically targets the core gene sequence of hepatitis B virus by synthesizing short interfering RNA (siRNA) in vivo, and to assess the inhibitory effect of this siRNA on HBV replication.


METHODS
The eukaryotic expression plasmid pHBV1.3, which contains 1.3-fold-overlength genome of HBV, were cotransfected into HepG2 cells with either the RNAi plasmid pSIHBV/C or unrelated control plasmid pSI. At 24, 48, 72 hours post transfection (p.t.), the levels of HBsAg and HBeAg in the cell culture medium were determined with Abbott MEIA Kits. The expression of intracellular viral proteins was also determined by immunofluorescence staining.


RESULTS
Successfully constructed expressing siRNA vector pSIHBV which targeted the core gene of Hepatitis B virus.The introduction of RNAi plasmid was shown to efficiently and specifically inhibit the synthesis of surface and e antigen of HBV by Abbot MEIA kits, with inhibitory rates at 92%, 85% peaking at 24 hours p.t. Immunofluorescence staining showed that intracellular synthesis of viral antigen was sharply reduced to nearly background levels when the ratio of pSIHBV/C and pHBV1.3 was at 1:20, whereas the control vector did not exhibit any inhibitory effect on the replication and expression of HBV.


CONCLUSIONS
Our results demonstrate that the short interfering RNA targeting HBV core gene exerts robust inhibition on HBV viral replication, suggesting that RNAi-based anti-HBV replication strategy may represent a potentially efficacious approach to the clinical management of HBV infection.
Tang
Ni
N
Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Huang
Ai-long
AL
Zhang
Bing-qiang
BQ
Yan
Ge
G
He
Tong-Chuan
TC
D005455:Fluorescent Antibody Technique; D015316:Genetic Therapy; D006509:Hepatitis B / Q000628:therapy*; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000502:physiology; D006801:Humans; D034622:RNA Interference; D014779:Virus Replication
D006512:Hepatitis B Core Antigens