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Published online ahead of print on 18 March 2009 as doi:10.1099/vir.0.010579-0
Journal of General Virology 2009;90:1812.

A more recent version of this article appeared on August 1, 2009 J Gen Virol (2009), DOI 10.1099/vir.0.010579-0
© 2009 Society for General Microbiology
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Targeted delivery of hepatitis C virus specific shRNA in mouse liver using Sendai virosomes

Nithya Subramanian1, Prashant Mani2, Swagata Roy1, Sivakumar Vadivel Gnanasundram1, Debi P Sarkar2 and Saumitra Das1,3

1 Indian Institute of Science;
2 University of Delhi, South Campus

3 E-mail: sdas{at}mcbl.iisc.ernet.in

Internal ribosome entry site (IRES) mediated translation of input viral RNA is the initial obligatory step for the replication of positive stranded genome of hepatitis C virus (HCV). Previously we have shown the importance of the GCAC sequence near initiator AUG within the stem and loop IV (SL-IV) region in mediating ribosome assembly on the HCV-RNA. Here, we demonstrate selective inhibition of HCV-IRES mediated translation using short-hairpin RNA (shRNA) targeting the same site within the HCV-IRES. The sh-SLIV showed significant inhibition of viral RNA replication in human hepatocellular carcinoma (Huh7) cell line harboring HCV monocistronic replicon. More importantly, co-transfection of infectious HCV-H77s RNA and sh-SLIV in Huh7.5 cells, successfully demonstrated significant decrease of viral RNA in HCV cell culture. Additionally, we report for the first time, the targeted delivery of sh-SLIV RNA into mice liver using Sendai virosome and demonstrate selective inhibition of HCV IRES mediated translation. Results provide the 'proof of concept' that Sendai virosome could be used for the efficient delivery of shRNAs into liver tissue to block HCV replication.

Received 26 January 2009; accepted 18 March 2009.




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