Comparative Analysis of Intracellular Trans-Splicing Ribozyme Activity Against Hepatitis C Virus Internal Ribosome Entry Site
Internal ribosome entry site (IRES) of the hepatitis C virus (HCV) is known to be essential for HCV replication and most conserved among HCV variants. Hence, IRES RNA is a good therapeutic target for RNA-based inhibitors, such as ribozymes. We previously proposed a new anti-HCV modulation strategy based on trans-splicing ribozymes, which can selectively replace HCV transcripts with a new RNA that exerts anti-HCV activity. To explore this procedure, sites which are accessible to ribozymes in HCV IRES were previously determined by employing an RNA mapping method in vitro. In this study, we evaluate the intracellular accessibility of the ribozymes by comparing the trans-splicing activities in cells of several ribozymes targeting different sites of the HCV IRES RNA. We assessed the intracellular activities of the ribozymes by monitoring their target-specific induction degree of both reporter gene activity and cytotoxin expression. The ribozyme capable of targeting the most accessible site identified by the mapping studies then harbored the most active trans-splicing activity in cells. These results suggest that the target sites predicted to be accessible are truly the most accessible in the cells, and thus, could be applied to the development of various RNA-based anti-HCV therapies.
- Honda, M., M.R. Beard, L.H. Ping, and S.M. Lemon. 1999. A phylogenetically conserved stem-loop structure at the 5' border of the internal ribosome entry site of hepatitis C virus is required for cap-independent viral translation. J. Virol. 73, 1165-1174
- Jones, J.T., S.-W. Lee, and B.A. Sullenger. 1996. Tagging ribozyme reaction sites to follow trans-splicing in mammalian cells. Nat. Med. 2, 643-648
- Kohler, U., B.G. Ayre, H.M. Goodman, and J. Haseloff. 1999. Trans-splicing ribozymes for targeted gene delivery. J. Mol. Biol. 185, 1935-1950
- Park Y.-H., H.-S. Jung, B.-S. Kwon, and S.-W. Lee. 2003. Replacement of thymidine phosphorylase RNA with group I intron of Tetrahymena thermophila by targeted trans-splicing. J. Microbiol. 41, 340-344
- Phylactou, L.A., C. Darrah, and M.A.J. Wood. 1998. Ribozymemediated trans-splicing of a trinucleotide repeat. Nat. Genet. 18, 378-381
- Ryu, K.-J., J.-H. Kim, and S.-W. Lee. 2003. Ribozyme-mediated selective induction of new gene activity in hepatitis C virus internal ribosome entry site-expressing cells by targeted transsplicing. Mol. Ther. 7, 386-395
- Byun, J., N. Lan, M. Long, and B.A. Sullenger. 2003. Efficient and specific repair of sickle beta-globin RNA by trans-splicing ribozymes. RNA 9, 1254-1263
- Watanabe, T. and B.A. Sullenger. 2000. Induction of wild-type p53 activity in human cancer cells by ribozymes that repair mutant p53 transcripts. Proc. Natl. Acad. Sci. USA 97, 8490-8494
- Shin, K.-S., B.A. Sullenger, and S.-W. Lee. 2004. Ribozyme-mediated induction of apoptosis in human cancer cells by targeted repair of mutant p53 RNA. Mol. Ther. 10, 365-372
- Lauer, G.M. and B.D. Waker. 2001. Hepatitis C virus infection. N. Engl. J. Med. 345, 41-52
- Ryu, K.-J. and S.-W. Lee. 2003. Identification of the most accessible sites to ribozymes on the hepatitis C virus internal ribosome entry site. J. Biochem. Mol. Biol. 36, 538-544
- Hugle, T. and A. Cerny. 2003. Current therapy and new molecular approaches to antiviral treatment and prevention of hepatitis C. Rev. Med. Virol. 13, 361-371
- Rogers, C.S., C.G. Vanoye, B.A. Sullenger and A.L. George, Jr. 2002. Functional repair of a mutant chloride channel using a trans-splicing ribozyme. J. Clin. Invest. 110, 1783-1798
Lan, N., R.P. Howrey, S.-W. Lee, C.A. Smith, and B.A. Sullenger. 1998. Ribozyme-mediated repair of sickle
$\beta$-globin mRNAs in erythrocyte precursors. Science 280, 1593-1596.
- Rosenberg, S. 2001. Recent advances in the molecular biology of hepatitis C virus. J. Mol. Biol. 313, 451-464
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