Abstract
Oligonucleotide analogs bearing modified phosphodiester linkages have been the focus of considerable interest in the antisense field, with methylphosphonates and phosphorothioates being the most extensively studied derivatives to date (1–3). These modifications, whereby a single nonbridging oxygen atom is replaced with either a methyl group or sulfur atom, generate certain very desirable oligomer characteristics (4–7), such as resistance toward nucleases, retention of the ability to form duplexes with natural DNA or RNA, and, relative to the phosphorothioate derivative, an ability to stimulate RNase H activity. Unfortunately, and in contrast to natural DNA, the phosphorus center is rendered chiral by these substitutions. Thus, for a deoxyoligonucleotide with n phosphodiester linkages, there can be 2n stereo-isomers with perhaps only one being the most active. Recent attempts to overcome this problem have focused on developing chemistries that either are stereoselective and generate oligomers enriched in one diastereomer (8,9) or are designed to synthesize new, phosphorus achiral analogs. In addition to being phosphorus achiral, perhaps new analogs may also have unique and attractive properties relative to the antisense area.
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References
Beaucage, S. L. and Iyer, R. P. (1992) Advances in the synthesis of oligonucleotides by the phosphoramidite approach. Tetrahedron 48, 2223–2311.
Englisch, U. and Gauss, D. H. (1991) Chemically modified oligonucleotides as probes and inhibitors. Angewandte Chemie Int. Edition 30, 613–722.
Uhlmann, E. and Peyman, A. (1990) Antisense oligonucleotides: a new therapeutic principle. Chem. Rev. 90, 544–584.
Goodchild, J. (1990) Conjugates of oligonucleotides and modified oligonucleotides: A review of their synthesis and properties. Bioconjugate Chemistry 2, 165–187.
Stein, C. A. (1989) Phosphorothioate oligodeoxynucleotide analogues, in Oligodeoxynucleotides. Antisense Inhibitors of Gene Expression (Cohen, J., ed.) Macmillan Press, London, pp. 97–117.
Ts’o, P. O. P., Miller, P. S., Aurelian, L., Murakami, A., Agris, C, Blake, K. R., Lin, S.-B., Lee, B. L., and Smith, C. C. (1987) An approach to chemotherapy based on base sequence information and nucleic acid chemistry, in Biological Approaches to the Controlled Delivery of Drugs. Annals of the New York Academy of Sciences, vol. 507, New York Academy of Sciences, New York, pp. 220–241.
Stein, C. A. and Cohen, J. S. (1988) Oligodeoxynucleotides as inhibitors of gene expression: a review. Cancer Res. 48, 2659–2668.
Lesnikowski, Z. J., Jaworska, M., and Stec, W. J. (1990) Octa(thymidine methanephosphonates) of partially defined stereochemistry: synthesis and effect of chirality at phosphorus on binding to pentadecadeoxyriboadenylic acid. Nucl. Acids Res. 18, 2109–2115.
Stec, W. J., Grajkowski, A., Koziolkiewicz, M., and Uznanski, B. (1991) Novel route to oligo(deoxyribonucleoside phosphorothioates) Stereocontrolled synthesis of P-chiral oligo(deoxyribonucleoside phosphorothioates). Nucl. Acids Res. 19, 5883–5888.
Brill, W. K.-D., Tang, J.-Y., Ma, Y.-X., and Caruthers, M. H. (1989) Synthesis of oligodeoxynucleoside phosphorodithioates via thioamidites. J. Am. Chem. Soc. 111, 2321,2322.
Nielsen, J., Brill, W. K.-D., and Caruthers, M. H. (1988) Synthesis and characterization of dinucleoside phosphorodithioates. Tetrahedron Lett. 29, 2911–2914.
Grandas, A., Marshall, W. S., Nielsen, J., and Caruthers, M. H. (1989) Synthesis of deoxycytidine oligomers containing phosphorodithioate linkages. Tetrahedron Lett. 30, 543–546.
Porritt, G. M. and Reese, C. B. (1989) Nucleoside phosphonodithioates as intermediates in the preparation of dinucleoside phosphorodithioates and phosphorothioates. Tetrahedron Lett. 30, 4713–4716.
Caruthers, M. H., Beaton, G., Cummins, L., Dellinger, D., Graff, D., Ma, Y.-X., Marshall, W. S., Sasmor, H., Shankland, P., Wu, J., and Yau, E. K. (1991) Chemical and biochemical studies with dithioate DNA. Nucleosides and Nucleotides 10, 47–59.
Caruthers, M. H., Beaton, G., Wu, J. V., and Wiesler, W. T. (1992) Methods in Enzymology, vol. 211 (Lilley, D. M. J. and Dahlberg, J. E., eds.) Academic Press, NY, pp. 3–20.
Dahl, O. (1991) Preparation of nucleoside phosphorothioates, phosphorodithioates and related compounds. Sulfur Reports, 11, and references cited therein, 167–192.
Beaton, G., Dellinger, D., Marshall, W. S., and Caruthers, M. H. (1991) Synthesis of oligonucleotide phosphorodithioates, in Oligonucleotides and Analogues, A Practical Approach (Eckstein, F., ed.) IRL Press, Oxford, pp. 109–135.
Brill, W. K.-D., Nielsen, J., and Caruthers, M. H. (1991) Synthesis of deoxydinucleoside phosphorodithioates. J. Am. Chem. Soc. 113, 3972–3980.
Caruthers, M. H., Barone, A. D., Beaucage, S. L., Dodds, D. R., Fisher, E. F., McBride, L. J., Matteucci, M. D., Stabinski, Z., and Tang, J.-Y. (1987) Chemical synthesis of deoxyoligonucleotides by the phosphoramidite method, in Methods in Enzymology, vol. 154 (Wu, R., ed.) Academic Press, NY, pp. 287–313.
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Wiesler, W.T., Marshall, W.S., Caruthers, M.H. (1993). Synthesis and Purification of Phosphorodithioate DNA. In: Agrawal, S. (eds) Protocols for Oligonucleotides and Analogs. Methods in Molecular Biology, vol 20. Humana Press. https://doi.org/10.1385/0-89603-281-7:191
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DOI: https://doi.org/10.1385/0-89603-281-7:191
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