RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells.
RNA interference represents an exciting new technology that could have therapeutic applications for the treatment of viral infections. Hepatitis C virus (HCV) is a major cause of chronic liver disease and affects >270 million individuals worldwide. The HCV genome is a single-stranded RNA that functions as both a messenger RNA and replication template, making it an attractive target for the study of RNA interference. Double-stranded small interfering RNA (siRNA) molecules designed to target the HCV genome were introduced through electroporation into a human hepatoma cell line (Huh-7) that contained an HCV subgenomic replicon. Two siRNAs dramatically reduced virus-specific protein expression and RNA synthesis to levels that were 90% less than those seen in cells treated with negative control siRNAs. These same siRNAs protected naive Huh-7 cells from challenge with HCV replicon RNA. Treatment of cells with synthetic siRNA was effective >72 h, but the duration of RNA interference could be extended beyond 3 weeks through stable expression of complementary strands of the interfering RNA by using a bicistronic expression vector. These results suggest that a gene-therapeutic approach with siRNA could ultimately be used to treat HCV.
Wilson
Joyce A
JA
Ontario Cancer Institute, 620 University Avenue, Suite 706, Toronto, ON, Canada M5G 2C1.
Jayasena
Sumedha
S
Khvorova
Anastasia
A
Sabatinos
Sarah
S
Rodrigue-Gervais
Ian Gaël
IG
Arya
Sudha
S
Sarangi
Farida
F
Harris-Brandts
Marees
M
Beaulieu
Sylvie
S
Richardson
Christopher D
CD
D000911:Antibodies, Monoclonal / Q000378:metabolism; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D018274:Electroporation; D005822:Genetic Vectors; D006526:Hepatitis C / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000166:cytology*; D008957:Models, Genetic; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D012313:RNA / Q000378:metabolism*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000502:physiology*; D012367:RNA, Viral / Q000235:genetics*; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D014779:Virus Replication / Q000235:genetics / Q000502:physiology*
D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D012313:RNA