title: Inhibition of translation of hepatitis C virus RNA by 2-modified antisense oligonucleotides.
abstract: 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.
authors.affiliation: Isis Pharmaceuticals, Carlsbad, CA 92008, USA.
authors.forename: V
authors.initials: V
authors.lastname: Brown-Driver
authors.forename: T
authors.initials: T
authors.lastname: Eto
authors.forename: E
authors.initials: E
authors.lastname: Lesnik
authors.forename: K P
authors.initials: KP
authors.lastname: Anderson
authors.forename: R C
authors.initials: RC
authors.lastname: Hanecak
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D016174:Hepacivirus / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000166:cytology; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects*; D012367:RNA, Viral / Q000235:genetics; D011817:Rabbits; D013873:Thionucleotides / Q000494:pharmacology; D014758:Viral Core Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; D020319:Oligodeoxyribonucleotides, Antisense; D016376:Oligonucleotides, Antisense; D012367:RNA, Viral; D013873:Thionucleotides; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus