Effective Inhibition of Hepatitis B Virus Replication by Small Interfering Rnas Expressed from Human Foamy Virus Vectors.
From: College of Life Sciences, Shaanxi Normal University, Xi’an, Shaanxi 710062, P.R. China.
International journal of molecular medicine
- Publish Date: Apr 2007
- ISSN: 1107-3756
- Volume: 19
- Issue: 4
- Pages: 705-11
- Medium: Print
- Language: English
- Citation (JAMA): Sun Yan, Li Zhi, Li Lijun, et al. Effective Inhibition of Hepatitis B Virus Replication by Small Interfering Rnas Expressed from Human Foamy Virus Vectors.. Int. J. Mol. Med. Apr 2007;19:705-11
Abstract
RNA interference (RNAi) mediated by double- stranded small interfering RNA (siRNA) is a novel mechanism of sequence-specific, post-transcriptional gene silencing. There has been much research into the use of RNAi for the treatment of human diseases. Many viruses, including hepatitis B virus (HBV), are susceptible to inhibition by this mechanism. However, for RNAi to be efficacious therapeutically, effective RNAi targeting sequences and a suitable delivery system are required. In this study, we employed a polymerase chain reaction (PCR)-based siRNA expression strategy to rapidly screen for effective siRNA sequences. Two effective siRNAs sequences (designated as S2 and X1) which reduced the HBV RNA by >90% were identified. For delivering the siRNAs, they were cloned into a human foamy virus (HFV)-based vector to generate single siRNA expression vectors HFVU6-siS2, HFVU6-siX1 and a dual siRNA expression vector HFVU6-siSX. The results showed that these siRNA vectors effectively inhibited multiple HBV gene expression and viral DNA replication based on ELISA and quantitative PCR analysis. HFVU6-siSX which simultaneously expressed two siRNAs that targeted the S and X genes of HBV is the most potent inhibitor of HBV replication. In addition, the repression of HBV RNA and DNA was stable for up to 3 months post-transduction as determined by RT-PCR and Southern blotting. Collectively, the PCR-based siRNA expression strategy provides a rapid and easy approach for testing candidate anti-HBV siRNA sequences and for cloning selected siRNA expression cassettes into a vector. RNAi based on the HFV vector was able to achieve effective, long-term inhibition of HBV gene expression and viral DNA replication. The combination of the two techniques may provide a powerful tool in the treatment of viral infection.
Mesh Headings (Keywords): Cells, Cultured, Gene Therapy, Genetic Vectors, Hepatitis B, Hepatitis B virus, Humans, RNA Interference, RNA, Small Interfering, Spumavirus, Viral Proteins, Virus Replication
Check for Full Text / PubMed Unique Identifier (PMID): 17334648
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