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Activity of Core‐Modified 10–23 DNAzymes against HCV
Author(s) -
Robaldo Laura,
BerzalHerranz Alfredo,
Montserrat Javier M.,
Iribarren Adolfo M.
Publication year - 2014
Publication title -
chemmedchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.817
H-Index - 100
eISSN - 1860-7187
pISSN - 1860-7179
DOI - 10.1002/cmdc.201402222
Subject(s) - internal ribosome entry site , deoxyribozyme , luciferase , translation (biology) , biology , microbiology and biotechnology , rna , untranslated region , chemistry , reticulocyte , virology , transfection , messenger rna , dna , cell culture , biochemistry , genetics , gene
The highly conserved untranslated regions of the hepatitis C virus (HCV) play a fundamental role in viral translation and replication and are therefore attractive targets for drug development. A set of modified DNAzymes carrying (2′ R )‐, (2′ S )‐2′‐deoxy‐2′‐ C ‐methyl‐ and ‐2′‐ O ‐methylnucleosides at various positions of the catalytic core were assayed against the 5′‐internal ribosome entry site element (5′‐IRES) region of HCV. Intracellular stability studies showed that the highest stabilization effects were obtained when the DNAzymes′ cores were jointly modified with 2′‐ C ‐methyl‐ and 2′‐ O ‐methylnucleosides, yielding an increase by up to fivefold in the total DNAzyme accumulation within the cell milieu within 48 h of transfection. Different regions of the HCV IRES were explored with unmodified 10–23 DNAzymes for accessibility. A subset of these positions was tested for DNAzyme activity using an HCV IRES‐firefly luciferase translation‐dependent RNA (IRES‐FLuc) transcript, in the rabbit reticulocyte lysate system and in the Huh‐7 human hepatocarcinoma cell line. Inhibition of IRES‐dependent translation by up to 65 % was observed for DNAzymes targeting its 285 position, and it was also shown that the modified DNAzymes are as active as the unmodified one.