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Utilisation de désoxyribozymes contre l'infection par le virus de l'hépatite C
| Content Provider | Semantic Scholar |
|---|---|
| Author | Trépanier, Janie |
| Copyright Year | 2008 |
| Abstract | Hepatitis C virus (HCV) infects over 123 million people worldwide. Chronically infected patients may eventually develop cirrhosis and hepatocellular carcinoma. Current therapy with interferon and ribavirin still fails to clear the virus and generates important side effects. The development of more unconventional HCV therapeutics is therefore required to identify novel agents capable of eliminating the virus and be better tolerated by patients. The overall aimof this project is to design deoxyribozymes, small sing~e-stranded DNA molecules with catalytic activity âgainst RNA targets, in order to cleave HCV RNA and stop its replication. The method used consisted in synthesizing deoxyribozymes to specifically cleave the HCV sequence encoding the highly conserved core protein. The cleavage efficiency of deoxyribozymes was first evaluated in an in vitro cell-fr~e assay, then ex vivo in cell culture and finally, in vivo in a mouse model. The in vitro and in vivo results indicate that the deoxyribozymes directed against the capsid RNA can cleave their target RNA in a specific and efficient manner. The most active deoxyribozyme was then modified by the addition of 2'-O-methyl adducts in order to improve its catalytic activity, increase its half-life and lower its potential toxicity. The efficiency of the modified molecule was then evaluated by transfecting hepatic cell lines containing genomic length HCV RNA. The real-time RT-PCR results indicated that 23.4 nM of this deoxyribozyme was sufficient to reduce the HCV target RNA by 63% ± 2.7% standard error of the mean after 24 h. This concentration was shown to be 40 times lower than the amount normally reported for this type of molecule. Moreover, there was a corresponding reduction in the viral capsid prote in levels as demonstrated by Western blot and immunofluorescence. The results obtained with our modified molecule demonstrate that it is one of the most effective deoxyribozyme ih cell culture. In conclusion, our results indicate that the use of· deoxyribozymes constitutes a promising therapeutic alternative for HCV infected patients. |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://papyrus.bib.umontreal.ca/xmlui/bitstream/handle/1866/6420/Trepanier_Janie_2008_these.pdf?isAllowed=y&sequence=1 |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
