Activity of core modified 10-23 DNAzymes against HCV

LAURA ROBALDO; ALFREDO BERZAL-HERRANZ; JAVIER MONTSERRAT; ADOLFO IRIBARREN

CHEMMEDCHEM

WILEY-V C H VERLAG GMBH

Año: 2014 vol. 9 p. 2271 - 2277

1860-7179

A set of modified DNAzymes carrying (2?R)-, (2?S)-2?-deoxy-2?-C-methyl and 2?-O-methyl nucleosides at different positions of the catalytic core have been assayed against the 5?-IRES region of HCV. Intracellular stability studies showed that the highest stabilization effects were obtained when the DNAzyme?s cores were jointly modified with 2?-C-methyl and 2?-O-methylnucleosides, yielding up to five fold increase in the total DNAzyme accumulation within the cell milieu after 48 hours of transfection.             Different regions of HCV IRES have been explored with unmodified 10-23 DNAzyme for accessibility. A subset of these positions has been tested for DNAzyme activity using a HCV IRES-Firefly luciferase translation dependent RNA (IRES-FLuc) transcript, in Rabbit Reticulocyte Lysate System and in Huh-7 human cells. Inhibition of IRES dependent translation up to 65% were obtained for DNAzymes targeting its 285 position, finding that the modified DNAzymes were as active as the unmodified one.