Site-specific cleavage of HCV genomic RNA and its cloned core and NS5B genes by DNAzyme.

摘要: Background and Aims:  The 9600 nt hepatitis C virus (HCV) genomic RNA has only one internal ribosome entry site (IRES) for translation to a single polyprotein. In search of nucleic acid-based antiviral agents, two 10-23 DNAzymes were designed cleave the in IRES dependent polymerase (RDRP/NS5B) regions prevent replication HCV RNA. Methods: In vitro cleavage by specific DNAzyme, CDz NS5B NDz was carried out using synthesized runoff transcripts core genes. Cleavage mRNAs DNAzyme (Dz) HepG2 cells assessed reverse transcription chain reaction (RT-PCR) from co-transfected with cloned or gene its respective DNAzyme. Suppression protein expression due mRNA Dz determined Western blot analysis fluorescence intensity fluorescent-tagged expressed protein. Reduction activity enzymatic assays. Results:  cleaved their generated transcripts. Both expressions genes reduced substantially when Dz. RDRP accompanied enzyme activity. Increased cleavage, inhibition expression, reduction observed on increasing concentration. Conclusion:  Core targeted can be used controlling RNA.