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Potent and selective inhibition of gene expression by an antisense heptanucleotide

Nature Biotechnology volume 14, pages 840–844 (1996)Cite this article

Abstract

Factors that govern the specificity of an antisense oligonucleotide (ON) for its target RNA include accessibility of the targeted RNA to ON binding, stability of ON/RNA complexes in cells, and susceptibility of the ON/RNA complex to RNase H cleavage. ON specificity is generally proposed to be dependent on its length. To date, virtually all previous antisense experiments have used 12–25 nt-long ONs. We explored the antisense activity and specificity of short (7 and 8 nt) ONs modified with C-5 propyne pyrimidines and phosphorothioate internucleotide linkages. Gene-selective, mismatch sensitive, and RNase H-dependent inhibition was observed for a heptanucleotide ON. We demonstrated that the flanking sequences of the target RNA are a major determinant of specificity. The use of shorter ONs as antisense agents has the distinct advantage of simplified synthesis. These results may lead to a general, cost-effective solution to the development of antisense ONs as therapeutic agents.

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References

  1. Stein, C.A. and Cheng, Y.-C. 1993. Antisense oligonucleotides as therapeutic agents—is the bullet really magical? Science 261: 1004–1012.

    Article  CAS  Google Scholar 

  2. Stein, C.A. and Krieg, A.M. 1994. Problems in interpretation of data derived from in vitro and in vivo use of antisense oligodeoxynucleotides. Antisense Research and Development 4: 67–69.

    Article  CAS  Google Scholar 

  3. Wagner, R.W. 1994. Gene inhibition using antisense oligodeoxynucleotides. Nature 372: 333–335.

    Article  CAS  Google Scholar 

  4. Woolf, T.M., Melton, D.A., and Jennings, C.G. 1992 Specificity of antisense oligonucleotides in vivo. Proc. Natl. Acad. Sci. USA 89: 7305–7309.

    Article  CAS  Google Scholar 

  5. Herschlag, D. 1991. Implications of ribozyme kinetics for targeting the cleavage of specific RNA molecules in vivo: more isn't always better. Proc. Natl. Acad. Sci. USA 88: 6921–6925.

    Article  CAS  Google Scholar 

  6. Wagner, R.W., Matteucci, M.D., Lewis, J.G., Gutierrez, A.J., Moulds, C., and Froehler, B.C. 1993. Antisense gene inhibition by oligonucleotides containing C-5 propyne pyrimidines. Science 260: 1510–1513.

    Article  CAS  Google Scholar 

  7. Rittner, K., Burmester, C., and Sczakiel, G. 1993. In vitro selection of fast-hybridizing and effective antisense RNAs directed against the human immunodeficiency virus type 1. Nucleic Acids Res. 21: 1381–1387.

    Article  CAS  Google Scholar 

  8. Lima, W.F., Monia, B.P., Ecker, D.J., and Freier, S.M. 1992 Implication of RNA structure on antisense oligonucleotide hybridization kinetics. Biochemistry 31: 12055–12061.

    Article  CAS  Google Scholar 

  9. Fenster, S.D., Wagner, R.W., Froehler, B.C., and Chin, D.J. 1994. Inhibition of Human Immunodeficiency Virus type-1 env expression by C-5 propyne oligonucleotides specific for Rev Response element stem loop V. Biochemistry 33: 8391–8398.

    Article  CAS  Google Scholar 

  10. Persson, C., Wagner, E.G.H., and Nordström, K. 1990. Control of replication of plasmid R1: structures and sequences of the antisense RNA, CopA, required for its binding to the target RNA, CopT. EMBO J. 9: 3767–3775.

    Article  CAS  Google Scholar 

  11. Mishra, R.K. and Toulme, J.J. 1994. In vitro selection of antisense oligonucleotides targeted to a hairpin structure. Comptes Rendus de L Academie des Sciences. Serie III, Sciences de la Vie 317: 977–982.

    CAS  PubMed  Google Scholar 

  12. Laptev, A.V., Lu, Z., Colige, A., and Prockop, D.J. 1994. Specific inhibition of expression of a human collagen gene (COL1A1) with modified antisense oligonucleotides. The most effective target sites are clustered in double-stranded regions of the predicted secondary structure for the mRNA. Biochemistry 33: 11033–11039.

    Article  CAS  Google Scholar 

  13. Morgan, R., Edge, M., and Colman, A. 1993 A more efficient and specific strategy in the ablation of mRNA in Xenopus laevis using mixtures of antisense oligos. Nucleic Acids Res. 21: 4615–4620.

    Article  CAS  Google Scholar 

  14. Homann, M., Rittner, K., and Sczakiel, G. 1993. Complementary large loops determine the rate of RNA duplex formation in vitro in the case of an effective antisense RNA directed against the human immunodeficiency virus type 1. J. Mol. Biol. 233: 7–15.

    Article  CAS  Google Scholar 

  15. Duroux, I., Godard, G., Boidot-Forget, M., Schwab, G., Hélène, C., and Saison-Behmoaras, T. 1995. Rational design of point mutation-selective antisense DNA targeted to codon 12 of Ha-ras mRNA in human cells. Nucleic Acids Res. 23: 3411–3418.

    Article  CAS  Google Scholar 

  16. Moulds, C., Lewis, J.G., Froehler, B.C., Grant, D., Huang, T., Milligan, J.F., Matteucci, M.D., and Wagner, R.W. 1995 Site and mechanism of antisense inhibition by C-5 propyne oligonucleotides. Biochemistry 34: 5044–5053.

    Article  CAS  Google Scholar 

  17. Lin, K.-Y., Pudlo, J.S., Jones, R.J., Bischofberger, N., Matteucci, M.D., and Froehler, B.C. 1994. Oligodeoxynucleotides containing 5-(1-propynyl)-2′-deoxyuridine formacetal and thioformacetal dimer synthons. Bioorganic & Medicinal Chem. Letters 4: 1061–1064.

    Article  CAS  Google Scholar 

  18. Gossen, M. and Bujard, H. 1992. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA 89: 5547–5551.

    Article  CAS  Google Scholar 

  19. Hanvey, J.C., Peffer, N.J., Bisi, J.E., Thomson, S.A., Cadilla, R., Josey, J.A., Hassman, C.F., Bonham, M.A., Au, K.G., Carter, S.G., Bruckenstein, D.A., Boyd, A.L., Noble, S.A., and Babiss, L.E. 1992. Antisense and antigene properties of peptide nucleic acids. Science 258: 1481–1485.

    Article  CAS  Google Scholar 

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  1. Richard W. Wagner: Corresponding author.

Authors and Affiliations

  1. Gilead Sciences, 353 Lakeside Dr., Foster City, CA, 94404

    Richard W. Wagner, Mark D. Matteucci, Deborah Grant, Teresa Huang & Brian C. Froehler

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  1. Richard W. Wagner
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  4. Teresa Huang
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Wagner, R., Matteucci, M., Grant, D. et al. Potent and selective inhibition of gene expression by an antisense heptanucleotide. Nat Biotechnol 14, 840–844 (1996). https://doi.org/10.1038/nbt0796-840

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