Inhibition of in vitro transcription by oligodeoxynucleotides.

Single-stranded short phosphodiester oligodeoxynucleotides have been used to inhibit in vitro T7 transcription system. These oligodeoxynucleotides were complementary to either the 3'-5' or 5'-3' strand of the transcription initiation site of a plasmid containing the gag region of HIV. Our results show that incubation of this plasmid DNA with the oligodeoxynucleotide complementary to the template DNA strand (3'-5', sense oligo) showed efficient inhibition of transcription. Incubation of this plasmid with the oligodeoxynucleotide complementary to the 5'-3' strand (antisense oligo) or a random oligodeoxynucleotide failed to do so. The inhibition of gag transcription was specific since the sense oligo failed to prevent transcription of a plasmid containing U2 RNA sequences. The inhibition of transcription was not limited to T7 RNA polymerase but was also observed with SP6 RNA polymerase.

[1]  P. Zamecnik,et al.  Electron micrographic studies of transport of oligodeoxynucleotides across eukaryotic cell membranes. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Y. Cheng,et al.  Antisense oligonucleotides as therapeutic agents--is the bullet really magical? , 1993, Science.

[3]  B. Sproat,et al.  Comparative evaluation of seven oligonucleotide analogues as potential antisense agents. , 1993, Journal of medicinal chemistry.

[4]  Y. Cheng,et al.  Phosphorothioate oligonucleotides are inhibitors of human DNA polymerases and RNase H: implications for antisense technology. , 1992, Molecular pharmacology.

[5]  T. Pederson,et al.  The U2 small nuclear ribonucleoprotein particle associates with nuclear factors in a pre-mRNA independent reaction. , 1991, The Journal of biological chemistry.

[6]  C. Hutchison,et al.  Cleavage of HIV-1 gag polyprotein synthesized in vitro: sequential cleavage by the viral protease. , 1989, AIDS research and human retroviruses.

[7]  J. Maizel,et al.  Inhibition of human immunodeficiency virus in early infected and chronically infected cells by antisense oligodeoxynucleotides and their phosphorothioate analogues. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[8]  T. Pederson,et al.  U2 small nuclear RNP assembly in vitro. , 1989, Nucleic acids research.

[9]  P. Zamecnik,et al.  Oligodeoxynucleoside phosphoramidates and phosphorothioates as inhibitors of human immunodeficiency virus. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Toulmé,et al.  Oligodeoxynucleotides covalently linked to intercalating agents: a new class of gene regulatory substances. , 1985, Biochimie.

[11]  S. Liebhaber,et al.  Translationally associated helix-destabilizing activity in rabbit reticulocyte lysate. , 1984, The Journal of biological chemistry.

[12]  S. Agrawal,et al.  Cellular uptake of oligodeoxynucleotide phosphorothioates and their analogs. , 1994, Antisense research and development.

[13]  D. Melton.  Injected anti-sense RNAs specifically block messenger RNA translation in vivo. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[14]  M. Stephenson,et al.  Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. , 1978, Proceedings of the National Academy of Sciences of the United States of America.