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- Title
Interfering with hepatitis C virus IRES activity using RNA molecules identified by a novel in vitro selection method.
- Authors
Romero-López, Cristina; Barroso-delJesus, Alicia; Puerta-Fernández, Elena; Berzal-Herranz, Alfredo
- Abstract
Hepatitis C virus (HCV) infection is one of the world's major health problems, and the identification of efficient HCV inhibitors is a major goal. Here we report the isolation of efficient anti-HCV internal ribosome entry site (IRES) RNA molecules identified by a new in vitro selection method. The newly developed procedure consists of two sequential steps that use distinct criteria for selection: selection for binding and selection for cleaving. The selection protocol was applied to a population of more than 10(15) variants of an anti-hepatitis C virus ribozyme covalently linked to an aptamer motif. The ribozyme was directed against positions 357 to 369 of the HCV IRES, and the cleavage substrate was a 691-nucleotide-long RNA fragment that comprises the entire HCV IRES domain. After six selection cycles, seven groups of RNA variants were identified. A representative of each group was tested for its capacity to inhibit IRES activity using in vitro translation assays. All selected RNAs promoted significant inhibition, some by as much as 95%.
- Publication
Biological chemistry, 2005, Vol 386, Issue 2, p183
- ISSN
1431-6730
- Publication type
Journal Article
- DOI
10.1515/BC.2005.023