title: Modified (PNA, 2'-O-methyl and phosphoramidate) anti-TAR antisense oligonucleotides as strong and specific inhibitors of in vitro HIV-1 reverse transcription.
abstract: Natural beta-phosphodiester 16mer and 15mer antisense oligonucleotides targeted against the HIV-1 and HIV-2 TAR RNAs respectively were previously described as sequence-specific inhibitors of in vitro retroviral reverse transcription. In this work, we tested chemically modified oligonucleotide analogues: alpha-phosphodiester, phosphorothioate, methylphosphonate, peptide nucleic acid or PNA, 2'- o -methyl and (N3'-P5') phosphoramidate versions of the 16mer anti-TAR oligonucleotide. PNA, 2'- O -methyl and (N3'-P5') phosphoramidate oligomers showed a strong inhibitory effect compared with the unmodified 16mer, with reverse transcription inhibition (IC50) values in the nanomolar range. The inhibition was sequence-specific, as scrambled and mismatched control oligonucleotides were not able to inhibit cDNA synthesis. No direct binding of the 2'- O -methyl, PNA or (N3'-P5') phosphoramidate anti-TAR oligonucleotides to the HIV-1 reverse transcriptase was observed. The higher T m obtained with 2'- O -methyl, (N3'-P5') phosphoramidate and PNA molecules concerning the annealing with the stem-loop structure of the TAR RNA, in comparison with the beta-phosphodiester oligonucleotides, is correlated with their high inhibitory effect on reverse transcription.
authors.affiliation: EP-630, CNRS-Université Victor Segalen Bordeaux 2, 1 rue Camille Saint-Saëns, 33077 Bordeaux cedex, France.
authors.forename: F
authors.initials: F
authors.lastname: Boulmé
authors.forename: F
authors.initials: F
authors.lastname: Freund
authors.forename: S
authors.initials: S
authors.lastname: Moreau
authors.forename: P E
authors.initials: PE
authors.lastname: Nielsen
authors.forename: S
authors.initials: S
authors.lastname: Gryaznov
authors.forename: J J
authors.initials: JJ
authors.lastname: Toulmé
authors.forename: S
authors.initials: S
authors.lastname: Litvak
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D016325:HIV Long Terminal Repeat / Q000187:drug effects*; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors*; D008969:Molecular Sequence Data; D009692:Nucleic Acid Heteroduplexes / Q000187:drug effects; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D009841:Oligonucleotides / Q000737:chemistry / Q000494:pharmacology; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000494:pharmacology*; D009843:Oligoribonucleotides / Q000494:pharmacology; D009943:Organophosphorus Compounds / Q000494:pharmacology; D020135:Peptide Nucleic Acids / Q000494:pharmacology*; D013873:Thionucleotides / Q000494:pharmacology*
chemical_list: D019380:Anti-HIV Agents; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D009843:Oligoribonucleotides; D009943:Organophosphorus Compounds; D020135:Peptide Nucleic Acids; D013873:Thionucleotides; C116746:phosphodiester alpha; C032627:methylphosphonic acid; D054303:HIV Reverse Transcriptase