[Inhibition of gene expression by antisense DNA].
We demonstrated that unmodified and modified (phosphorothioate) oligonucleotides prevent cDNA synthesis by AMV, MMLV, or HIV reverse transcriptases. Antisense oligonucleotide/RNA hybrids specifically arrest primer extension. The blockage involves the degradation of the RNA fragment, bound to the antisense oligonucleotide, by reverse transcriptase associated RNase H activity. However, the phosphorothioate oligomer inhibited polymerization, by binding to the AMV- and MMLV-RTs, rather than to the template RNA, whereas there was no competitive binding of the phosphorothioate oligomer on the HIV RT during reverse transcription. Furthermore, the RNase H activity of HIV-RT was only slightly affected by the phosphorothioate oligonucleotide. The anti-HIV activities of phosphorothioate- or 5'-linked lipid-oligonucleotides are also described and some of the problems that still need to be solved are pointed out.
Takaku
H
H
Department of Industrial Chemistry, Chiba Institute of Technology.
Kim
S G
SG
Hatta
T
T
Takai
K
K
D009189:Avian Myeloblastosis Virus / Q000235:genetics; D001483:Base Sequence; D016373:DNA, Antisense / Q000502:physiology*; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D006678:HIV / Q000235:genetics; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics; D012194:RNA-Directed DNA Polymerase / Q000502:physiology
D016373:DNA, Antisense; D012194:RNA-Directed DNA Polymerase