Inhibition of translation of hepatitis C virus RNA by 2-modified antisense oligonucleotides

• Published in: Antisense & nucleic acid drug development Vol. 9; no. 2; p. 145
• Main Authors: Brown-Driver, V, Eto, T, Lesnik, E, Anderson, K P, Hanecak, R C
• Format: Journal Article
• Language: English
• Published: United States 01.04.1999
• Subjects: 5' Untranslated Regions; Animals; Hepacivirus - genetics; Humans; Liver - cytology; Oligodeoxyribonucleotides, Antisense - pharmacology; Oligonucleotides, Antisense - pharmacology; Protein Biosynthesis - drug effects; Rabbits; RNA, Viral - genetics; Thionucleotides - pharmacology; Viral Core Proteins - biosynthesis; Viral Core Proteins - genetics
• ISSN: 1087-2906
• DOI: 10.1089/oli.1.1999.9.145

Inhibition of hepatitis C virus (HCV) gene expression by antisense oligonucleotides was investigated using both a rabbit reticulocyte lysate in vitro translation assay and a transformed human hepatocyte cell expression assay. Screening of overlapping oligonucleotides complementary to the HCV 5' noncoding region and the core open reading frame (ORF) identified a region susceptible to translation inhibition between nucleotides 335 and 379. Comparison of 2'-deoxy-, 2'-O-methyl-, 2'-O-methoxyethyl-, 2'-O-propyl-, and 2'-fluoro-modified phosphodiester oligoribonucleotides demonstrated that increased translation inhibition correlated with both increased binding affinity and nuclease stability. In cell culture assays, 2'-O-methoxyethyl-modified oligonucleotides inhibited HCV core protein synthesis with comparable potency to phosphorothioate oligodeoxynucleotides. Inhibition of HCV core protein expression by 2'-modified oligonucleotides occurred by an RNase H-independent translational arrest mechanism.