Inhibition of translation of hepatitis C virus RNA by 2-modified antisense oligonucleotides.
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.
Brown-Driver
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Isis Pharmaceuticals, Carlsbad, CA 92008, USA.
Eto
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Lesnik
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Anderson
K P
KP
Hanecak
R C
RC
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
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