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The effect of antisense oligodeoxyribonucleotide (ODN) and antisense RNA on the simian virus 40 (SV40) DNA replication in a cell-free system was examined. The cell-free replication was performed with plasmid containing SV40 origin of DNA replication, T antigen and HeLa extract. DNA primase initiation sites in the SV40 origin of the replication were selected as target sites of antisense ODN and antisense RNA. 3 antisense ODNs [16-mer, 17-mer and 19-mer (random)] and 5 antisense RNAs [I (18 nts), II (19 nts), BS (58 nts, containin I), SN (123 nts, containing II) and G (60 nts, random)] were used to examine the effect on the replication. It was obseved that the more the concentration of antisense construct increases, the more DNA replication was inhibited. It was also shown that DNA replication was inhibited nonspecifically by random sequences (19-mer and G) and the effect of random RNA was larger than random ODN. In addition, antisense constructs on SV40 DNA replication were examined. Although the effect of antisense construct on SV40 DNA replication in vitro was not the same as in vivo (previous work), it was shown that SV40 DNA replication was inhibited to some extent by the antisense construct in the in vitro condition.
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