EFFECTS OF ANTISENSE OLIGODEOXYRIBONUCLEOTIDES ON THE INITIATION OF SIMIAN-VIRUS-40 DNA-REPLICATION

Two groups of primase initiation sites around the simian virus 40 (SV40) origin of replication were selected for studies on the effects on initiation of SV40 DNA replication by antisense oligodeoxyribonucleotides (ODNs), 16-mer and 17-mer. Each or both ODNs were treated to the culture of the African green monkey kidney cells producing T antigen (COS) transfected with a recombinant plasmid containing the SV40 origin of replication, pATSV-W. The replication efficiency of plasmid pATSV-W in COS was measured by the following methods: the resistance to digestion by the restriction enzyme, DpnI, the tritium-thymidine incorporation, and density labeling by bromodeoxyuridine incorporation. The results of replication analysis showed that SV40 DNA replication was inhibited to 63% by antisense ODNs. These results were confirmed by in vivo RNA priming assay.