Antiviral therapy: inhibition of Hepatitis C Virus expression by RNA interference directed against the NS5B region of the viral genome.
BACKGROUND
Hepatitis C virus (HCV) is a major public health problem with 170 million chronically infected people throughout the world. Currently, the only treatment available consists of a combination of pegylated interferon (INF-alpha) and ribavirin, but only half of the patients treated show a sufficient antiviral response. Thus there is a great need for the development of new treatments for HCV infections. RNA interference (RNAi) represents a new promising approach to develop effective antiviral drugs and has been extremely effective against HCV gene expression in short-term cell culture. Our aim was to determine the effect of RNAi directed against the NS5B-HCV region on HCV expression in a human hepatoma cell line that expresses HCV-subgenomic replicon (Huh7 HCV replicon cells).


METHODS
We transfected Huh7 HCV replicon cells with different concentrations of RNAi (100-200 nM) targeting the NS5B region of the viral genome. 2-6 days post-transfection HCV-RNA was quantified by semiquantitative and real-time RT-PCR, and HCV NS5B protein levels were assayed by western blot. Cell viability was also quantified by MTT assay.


RESULTS
Our results indicate that the NS5B-siRNAs used in this study can specifically inhibit HCV-RNA replication and protein expression (more than 90%) compared to control cells.


CONCLUSIONS
Synthetic siRNA against NS5BHCV inhibited HCV replication and viral proteins levels and thereby becomes a powerful strategy to combat hepatitis C virus.
Trejo-Avila
Laura
L
Laboratorio de Inmunología y Virología, Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Monterrey, Nuevo León, México.
Elizondo-González
Regina
R
Trujillo-Murillo
Karina Del C
Kdel C
Zapata-Benavides
Pablo
P
Rodríguez-Padilla
Cristina
C
Rivas-Estilla
Ana María
AM
D000998:Antiviral Agents / Q000494:pharmacology / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000187:drug effects / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D016679:Genome, Viral / Q000187:drug effects / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism; D006526:Hepatitis C / Q000188:drug therapy* / Q000378:metabolism; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology / Q000627:therapeutic use*; D012367:RNA, Viral / Q000187:drug effects / Q000235:genetics; D012093:Replicon / Q000187:drug effects; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000235:genetics* / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase