Abstract
Duplex formation between an oligonucleotide and a strand of mRNA can effectively inhibit the expression of a specific gene via interfering with the cellular protein synthesis process. The antisense technology (1) is regarded as a powerful tool in molecular biology. Advances in the field have also led to the development of many pharmaceutical/therapeutic applications, with a number of oligonucleotides undergoing human clinical trials at present (2). Making the best use of this approach requires a good understanding of the subject, careful planning, a knowledge of limitations and pitfalls, and a capacity to go beyond the basic antisense techniques.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wagner, R. W. (1994) Gene inhibition using antisense oligonucleotides. Nature 372, 333–335.
Akhtar, S. and Agrawal, S. (1997) In vivo studies with antisense oligonucleotides. Trends Pharmaceut. Sci. 18, 12–18.
Hallbook, F., Sahlen, A., and Catsicas, S. (1997) Characterization and evaluation of NGF antisense oligonucleotides: inhibition of NGF synthesis in transfected COS cells. Antisense Nucleic Acid Drug Dev. 7, 89–100.
Staeker, H., Van de Water, T. R., Lefebvre, P. P., Liu, W., Moghadassi, M., Galinovic-Schwartz, V., et al. (dy1996) NGF, BDNF and NT-3 play unique roles in the in vitro development and patterning of innervation of the mammalian innerear. Dev. Brain Res. 92, 49–60.
Taglialatela, G., Hibbert, C. J., Hutton, L. A., Werrbach-Perez, K., and Perez-Polo, J. R. (1996) Suppression of p140 (trkA) does not abolish NGF mediated recue of serum free PC12 cells. J. Neurochem. 66, 1826–1835.
Rickman, D. W. and Rickman, C. B. (1996) Suppression of trkB expression by antisense oligonucleotides alters a neuronal phenotype in the rod pathway of developing rat retina. Proc. Natl. Acad. Sci. USA 93, 12,564–12,569.
Barrett, G. L. and Bartlett, P. F. (1994) The p75 growth factor receptor mediates survival or death depending on the stage of sensory neuron development. Proc. Natl. Acad. Sci. USA 91, 6501–6505.
Stein, C. A. (1996) Exploiting the potential of antisense: beyond phosphorothioate oligodeoxynucleotides. Chem. Biol. 3, 319–323.
Tonkinson, J. L. and Stein, C. A. (1994) Patterns of intracellular compartmentalization,trafficking and acidification of 5’-fluorescein labeled phosphodiester and phosphorothioate oligodeoxynucleotides in HL60 cells. Nucleic Acids Res. 22, 4268–4275.
Still, W. C., Kahn, M., and Mitra, A. (1978) Rapid chromatographic technique for preperative seperations with moderate resolution. J. Org. Chem. 43, 2923–2925.
Iversen, P. L., Zhu, S., Meyer, A., and Zon, G. (1992) Cellular uptake and subcellular distribution of phosphorothioate oligonucleotides into cultured cells. Antisense Res. Dev. 2, 211–222.
Temsamami, J., Kubert, M., Tang, J., Padmapriya, A., and Agrawal, S. (1994) Cellular uptake of oligoeoxynucleotide phosphorothoates and their analogs. Antisense Res. Dev. 4, 35–42.
Goodchild, J. (1989) Inhibition of gene expression by oligonucleotides, in Oligodeoxynucleotides: Antisense Inhibitors of Gene Expression (Cohen, J. S.,ed.), CRC, Boca Raton, FL, pp. 53–78.
Beaucage, S. L. (1993) Oligodeoxyribonucleotide synthesis: phosphoramidite approach, in Protocols for Oligonucleotides and Analogs (Agrawal, S., ed.), Humana, Totowa, NJ, pp. 233–264.
Iden, C. R., Rieger, R. A., Torres, C., and Martin, L. B. (1996) Application of electrospray ionization mass spectrometry to the analysis of oligodeoxynucleotides, in Biochemical and Biotechnological Applications of Electrospray Ionization Mass Spectroscopy (Snyder, A. P., ed.), American Chemical Society, Washington, DC, pp. 281–293.
Donnis-Keller, H. (1979) Site specific enzymatic cleavage of RNA. Nucleic Acid Res. 7, 179–189.
Agrawal, S., Jiang, Z., Zhoa, Q., Shaw, D., Cai, Q., Roskey, A., et al. (dy1997) Mixed-backbone oligonucleotides as second generation antisense oligonucleotides:in vitro and in vivo studies. Proc. Natl. Acad. Sci. USA 94, 2620–2625.
Dean, N. M. and Griffey, R. H. (1997) Identification and characterization of second generation antisense oligonucleotides. Antisense Nucleic Acid Drug Dev. 7, 229–233.
Leonetti, J-P., Degols, G., and Lableu, B. (1990), Biological activity of oligonucleotide-poly (l) lysine conjugates: mechanism of cell uptake. Bioconjugate Chem. 1, 149–153.
Felgner, P. L., Gadek, T. R., Holm, M., Roman, R., Chan, H. W., Wenz, M., et al. (dy1989) Lipofection: a highly efficient lipid mediated DNA transfection procedure. Proc. Natl. Acad. Sci. USA 84, 7413–7417.
Van der Woude, I., Wagennar, A., Meekel, A. A. P., ter Beest, M. B. A., Ruiters, M. H. J., Engberts, J. B. F. N., et al. (dy1997) Novel pyridinium surfactants for efficient nontoxic in vitro gene delivery. Proc. Natl. Acad. Sci. USA 94, 1160–1165.
MacKellar, C., Graham, D., Will, D. W., Burgess, S., and Brown, T. (1992) Synthesis and physical properties of anti-HIV antisense oligonucleotides bearing terminal lipophilic groups. Nucleic Acids Res. 20, 3411–3417.
Gryzanov, S. M. and Lloyd, D. H. (1993) Modulation of oligonucleotide duplex and triplex stability via hydrophobic interactions. Nucleic Acids Res. 21, 5909–5915.
Atkinson, T. and Smith, M. (1984) Solid-phase synthesis of oligodeoxynucleotides by the phosphite triester method, in Oligonucleotide Synthesis:A Practical Approach (Gait, M. J., ed.), IRL, Oxford, pp. 35–81.
Zhao, Q., Temsamani, J., and Agrawal, S. (1995) Use of cyclodextrin and its derivatives as carriers for oligonucleotide delivery. Antisense Res. Dev. 5, 185–192.
Habus, I., Zhao, Q., and Agrawal, S. (1995) Synthesis, hybridization properties nuclease stability and cellular uptake of the oligonucleotide-amino-β-cyclodextrins and adamantane conjugates. Bioconjugate Chem. 6, 327–331.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, p. 7.39.
Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 3, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, p. 18.60.
Stephenson, M. L. and Zamecnik, P. C. (1978) Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide. Proc. Natl. Acad. Sci. USA 75, 285–288.
Leonetti, J. P., Rayner, B., Lemaitre, M., Gagnor, C., Milhaud, P. G., Imbach, J-L., et al. (dy1988) Antiviral activity of conjugates between poly(L-lysine) and synthetic oligodeoxyribonucleotides. Gene 72, 323–332.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Epa, W.R., Barrett, G.L., Bartlett, P.F. (2001). Oligonucleotides as Inhibitors of Protein Synthesis. In: Rush, R.A. (eds) Neurotrophin Protocols. Methods in Molecular Biology™, vol 169. Humana Press. https://doi.org/10.1385/1-59259-060-8:223
Download citation
DOI: https://doi.org/10.1385/1-59259-060-8:223
Publisher Name: Humana Press
Print ISBN: 978-0-89603-699-4
Online ISBN: 978-1-59259-060-5
eBook Packages: Springer Protocols