Inhibition of intracellular hepatitis C virus replication by synthetic and vector-derived small interfering RNAs.
Small interfering RNAs (siRNAs) efficiently inhibit gene expression by RNA interference. Here, we report efficient inhibition, by both synthetic and vector-derived siRNAs, of hepatitis C virus (HCV) replication, as well as viral protein synthesis, using an HCV replicon system. The siRNAs were designed to target the 5' untranslated region (5' UTR) of the HCV genome, which has an internal ribosomal entry site for the translation of the entire viral polyprotein. Moreover, the 5' UTR is the most conserved region in the HCV genome, making it an ideal target for siRNAs. Importantly, we have identified an effective site in the 5' UTR at which approximately 80% suppression of HCV replication was achieved with concentrations of siRNA as low as 2.5 nM. Furthermore, DNA-based vectors expressing siRNA against HCV were also effective, which might allow the efficient delivery of RNAi into hepatocytes in vivo using viral vectors. Our results support the feasibility of using siRNA-based gene therapy to inhibit HCV replication, which may prove to be valuable in the treatment of hepatitis C.
Yokota
Takanori
T
Department of Neurology and Neurological Science, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo 113-8519, Japan. tak-yokota.nuro@tmd.ac.jp
Sakamoto
Naoya
N
Enomoto
Nobuyuki
N
Tanabe
Yoko
Y
Miyagishi
Makoto
M
Maekawa
Shinya
S
Yi
Li
L
Kurosaki
Masayuki
M
Taira
Kazunari
K
Watanabe
Mamoru
M
Mizusawa
Hidehiro
H
D020121:5' Untranslated Regions; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D017930:Genes, Reporter; D016679:Genome, Viral; D016174:Hepacivirus / Q000378:metabolism*; D006526:Hepatitis C / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000737:chemistry / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D014779:Virus Replication
D020121:5' Untranslated Regions; D009841:Oligonucleotides; D034741:RNA, Small Interfering; D004247:DNA; D008156:Luciferases