Antisense Gene Inhibition by Oligonucleotides Containing C-5 Propyne Pyrimidines

• Published in: Science (American Association for the Advancement of Science) Vol. 260; no. 5113; pp. 1510 - 1513
• Main Authors: Wagner, Richard W., Matteucci, Mark D., Lewis, Jason G., Gutierrez, Arnold J., Moulds, Courtney, Froehler, Brian C.
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for the Advancement of Science 04-06-1993; American Association for the Advancement of Science
• Subjects: Alkynes - pharmacology; Animals; Base Sequence; Biological and medical sciences; Cell Line; Cell lines; Cell nucleus; Cercopithecus aethiops; Fluorescence; Fundamental and applied biological sciences. Psychology; Gene expression; Genetic regulation; HeLa cells; Humans; Inhibitory concentration 50; Microinjections; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Oligonucleotides; Oligonucleotides, Antisense - pharmacokinetics; Oligonucleotides, Antisense - pharmacology; Physiological aspects; Plasmids; Pyrimidine Nucleotides - pharmacokinetics; Pyrimidine Nucleotides - pharmacology; Rats; RNA; RNA - drug effects; Start codon; Thionucleotides - pharmacokinetics; Thionucleotides - pharmacology; Sequence specificity; RNA; Monkey; Metabolic inhibitor; Molecular targeting; Gene expression; Kidney; Dose activity relation; Vertebrata; Mammalia; Cell line; Structure activity relation; Primates
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.7684856

Phosphorothioate oligodeoxynucleotides containing the C-5 propyne analogs of uridine and cytidine bind RNA with high affinity and are potent antisense inhibitors of gene expression. In a cellular assay, gene-specific antisense inhibition occurred at nanomolar concentrations of oligonucleotide, was dose-dependent and exquisitely sensitive to sequence mismatches, and was correlated with the melting temperature and length of oligonucleotide. Activity was independent of RNA target site and cell type but was detectable only when the oligonucleotides were microinjected or delivered with cell-permeabilizing agents. These oligonucleotides may have important applications in therapy and in studies of gene function.