Targeted delivery of hepatitis C virus-specific short hairpin RNA in mouse liver using Sendai virosomes

• Published in: Journal of general virology Vol. 90; no. 8; pp. 1812 - 1819
• Main Authors: Subramanian, Nithya, Mani, Prashant, Roy, Swagata, Gnanasundram, Sivakumar Vadivel, Sarkar, Debi P, Das, Saumitra
• Format: Journal Article
• Language: English
• Published: Reading Soc General Microbiol 01-08-2009; Society for General Microbiology
• Subjects: Animals; Antiviral Agents - administration & dosage; Biological and medical sciences; Cell culture; Cell Line, Tumor; Fundamental and applied biological sciences. Psychology; Genomes; Hepacivirus - drug effects; Hepatitis C; Hepatitis C - drug therapy; Hepatitis C virus; Hepatocellular carcinoma; Humans; Internal ribosome entry site; Liver - virology; Luciferases - metabolism; Male; Mice; Microbiology; Miscellaneous; Nucleotide sequence; Replication; Ribosomes; RNA; RNA, Small Interfering - administration & dosage; RNA, Viral - genetics; RNA, Viral - metabolism; Sendai virus - genetics; Translation; Tumor cell lines; Viral Plaque Assay; Virology; Virosomes; Virus Replication - drug effects; RNA interference; Microbiology; Targeted therapy; Liver; Rodentia; Virosome; Virus; Gene silencing; Vertebrata; Mammalia; Mouse; Animal; Flaviviridae; Hepatitis C virus; Short hairpin RNA; Hepacivirus
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/vir.0.010579-0

1 Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore 560012, India
2 Department of Biochemistry, University of Delhi, South Campus, New Delhi 110021, India
Correspondence Saumitra Das sdas{at}mcbl.iisc.ernet.in
Internal ribosome entry site (IRES)-mediated translation of input viral RNA is the initial required step for the replication of the positive-stranded genome of hepatitis C virus (HCV). We have shown previously the importance of the GCAC sequence near the initiator AUG within the stem and loop IV (SLIV) region in mediating ribosome assembly on HCV RNA. Here, we demonstrate selective inhibition of HCV-IRES-mediated translation using short hairpin (sh)RNA targeting the same site within the HCV IRES. sh-SLIV showed significant inhibition of viral RNA replication in a human hepatocellular carcinoma (Huh7) cell line harbouring a HCV monocistronic replicon. More importantly, co-transfection of infectious HCV–H77s RNA and sh-SLIV in Huh7.5 cells successfully demonstrated a significant decrease in 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 virosomes and demonstrate selective inhibition of HCV-IRES-mediated translation. Results provide the proof of concept that Sendai virosomes could be used for the efficient delivery of shRNAs into liver tissue to block HCV replication.
These authors contributed equally to this work.
Two supplementary figures are available with the online version of this paper.