Abstract
Small interfering RNAs (siRNAs) and short hairpin RNAs (shRNAs) have been reported to suppress gene expression significantly. HCV seems a suitable candidate for targets of siRNAs, as HCV is a positive single-strand RNA virus and replicates in the cytoplasm. Efficient inhibition by siRNAs requires access to target RNAs, which usually possess secondary structure. We have shown that shRNAs suppressing the HCV internal ribosomal entry site (IRES) can inhibit different HCV genotypes grown in cell culture and replicon replication, suggesting the potential of siRNA as an additional therapeutic option against HCV infection.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Elbashir, S. M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K., and Tuschl, T. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411, 494–498.
Liu, J., Valencia-Sanchez, M. A., Hannon, G. J., and Parker, R. (2005) MicroRNA-dependent localization of targeted mRNAs to mammalian P-bodies. Nat. Cell Biol. 7, 719–723.
McCaffrey, A. P., Meuse, L., Pham, T. T., Conklin, D. S., Hannon, G. J., and Kay, M. A. (2002) RNA interference in adult mice. Nature 418, 38–39.
Kumar, M. and Carmichael, G. G. (1998) Antisense RNA: function and fate of duplex RNA in cells of higher eukaryotes. Microbiol. Mol. Biol. Rev. 62, 1415–1434.
Stark, G. R., Kerr, I. M., Williams, B. R., Silverman, R. H., and Schreiber, R. D. (1998) How cells respond to interferons. Annu. Rev. Biochem. 67, 227–264.
Uprichard, S. L., Boyd, B., Althage, A., and Chisari, F. V. (2005) Clearance of hepatitis B virus from the liver of transgenic mice by short hairpin RNAs. Proc. Natl. Acad. Sci. USA 102, 773–778.
Brummelkamp, T. R., Bernards, R., and Agami, R. (2002) A system for stable expression of short interfering RNAs in mammalian cells. Science 296, 550–553.
Barik, S. (2006) RNAi in moderation. Nat. Biotechnol. 24, 796–797.
Petersen, C. P., Bordeleau, M. E., Pelletier, J., and Sharp, P. A. (2006) Short RNAs repress translation after initiation in mammalian cells. Mol. Cell 21, 533–542.
Sen, A., Steele, R., Ghosh, A. K., Basu, A., Ray, R., and Ray, R. B. (2003) Inhibition of hepatitis C virus protein expression by RNA interference. Virus Res. 96, 27–35.
Kapadia, S. B., Brideau-Andersen, A, and Chisari, F. V. (2003) Interference of hepatitis C virus RNA replication by short interfering RNAs. Proc. Natl. Acad. Sci. USA 100, 2014–2018.
Randall, G., Grakoui, A., and Rice, C. M. (2003) Clearance of replicating hepatitis C virus replicon RNAs in cell culture by small interfering RNAs. Proc. Natl. Acad. Sci. USA 100, 235–240.
Seo, M. Y., Abrignani, S., Houghton, M., and Han, J. H. (2003) Small interfering RNA-mediated inhibition of hepatitis C virus replication in the human hepatoma cell line Huh-7. J. Virol. 77, 810–812.
Wilson, J. A., Jayasena, S., Khvorova, A., Sabatinos, S., Rodrigue-Gervais, I. G., Arya, S., et al. (2003) RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells. Proc. Natl. Acad. Sci. USA 100, 2783–2788.
Wilson, J. A. and Richardson, C. D. (2005) Hepatitis C virus replicons escape RNA interference induced by a short interfering RNA directed against the NS5b coding region. J. Virol. 79, 7050–7058.
Kanda, T., Steele, R., Ray, R., and Ray, R. B. (2007) Small interfering RNA targeted to hepatitis C virus 5′nontranslated region exerts potent antiviral effect. J. Virol. 81, 669–679.
Ray, R. B., Meyer, K., and Ray, R. (2000) Hepatitis C virus core protein promotes immortalization of primary human hepatocytes. Virology 271, 197–204.
Kanda, T., Basu, A., Steele, R., Wakita, T., Ryerse, J. S., Ray, R., and Ray, R. B. (2006) Generation of infectious hepatitis C virus in immortalized human hepatocytes. J. Virol. 80, 4633–4639.
Blight, K. J., McKeating, J. A., and Rice, C. M. (2002) Highly permissive cell lines for subgenomic and genomic hepatitis C virus RNA replication. J. Virol. 76, 13001–13014.
Lohmann, V., Korner, F., Koch, J., Herian, U., Theilmann, L., and Bartenschlager, R. (1999) Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell lines. Science 285, 110–113.
Pietschmann, T., Lohmann, V., Kaul, A., Krieger, N., Rinck, G., Rutter, G., et al. (2002) Persistent and transient replication of full-length hepatitis C virus genomes in cell culture. J. Virol. 76, 4008–4021.
Korf, M., Jarczak, D., Berger, C., Manns, M. P., and Kruger, M. (2005) Inhibition of hepatitis C virus translation and subgenomic replication by siRNAs directed against highly conserved HCV sequence and cellular HCV cofactors. J. Hepatol. 43, 225–234.
Acknowledgements
The authors would like to thank Drs. M. Kruger, R. Bartenschlager, T. Wakita, and C. M. Rice for providing reagents and Robert Steele for critical reading. This work was supported by research grant AI45144 from the National Institutes of Health.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Ray, R.B., Kanda, T. (2009). Inhibition of HCV Replication by Small Interfering RNA. In: Tang, H. (eds) Hepatitis C. Methods in Molecular Biology™, vol 510. Humana Press. https://doi.org/10.1007/978-1-59745-394-3_19
Download citation
DOI: https://doi.org/10.1007/978-1-59745-394-3_19
Publisher Name: Humana Press
Print ISBN: 978-1-58829-970-3
Online ISBN: 978-1-59745-394-3
eBook Packages: Springer Protocols