Antisense Oligonucleotides Against Inducible Nitric Oxide Synthase Reduce NO Production in RAW 264.7 Cells

Polish - Japan Issue ; Several oligodeoxyribonucleotides in a nuclease-resistant phosphorothioate form (S-ODNs)targeted to iNOS mRNA were tested in RAW 264.7 cells as potential antisense inhibitors.Antisense S-ODNs inhibited iNOS activity in a time- and dose-dependent manner. Maximalinhibitory effect was >90% while control S-ODNs were ineffective. Application of Lipofectin Reagent, which itself had no significant effect, greatly increased antisense activity. Decreased levelsof the target mRNA, as demonstrated by reverse transcriptase-polymerase chain reaction (RTPCR), suggest the RNase H mediated mechanism. ; Zeszyt specjalny. Polish - Japan Issue ; Several oligodeoxyribonucleotides in a nuclease-resistant phosphorothioate form (S-ODNs)targeted to iNOS mRNA were tested in RAW 264.7 cells as potential antisense inhibitors.Antisense S-ODNs inhibited iNOS activity in a time- and dose-dependent manner. Maximalinhibitory effect was >90% while control S-ODNs were ineffective. Application of Lipofectin Reagent, which itself had no significant effect, greatly increased antisense activity. Decreased levelsof the target mRNA, as demonstrated by reverse transcriptase-polymerase chain reaction (RTPCR), suggest the RNase H mediated mechanism.