Abstract
Solid-phase oligodeoxynucleotide synthesis has become a routine procedure in most molecular-biology laboratories. The reagents for the synthesis of unmodified oligomers have been available for several years, and novel commercially available reagents that permit the introduction of useful analogs into DNA are offered with increasing frequency. Solid-phase RNA synthesis is also becoming accessible to the molecular-biology community. If a suitable protected derivative is available, solid-phase synthesis is usually the preferred method of incorporating a nonstandard residue into an oligomer. However, there are still situations in which it is advantageous, or even essential, to approach the synthesis of modified oligonucleotides in a different way, namely by derivatizing unprotected oligonucleotides or their analogs.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Knorre, D. G. and Vlassov, V. V. (1985) Complementary-addressed (sequence-specific) modification of nucleic acids, in Progress in Nucleic Acid Research and Molecular Biology, vol. 32 (Cohn, W. E. and Moldave, K., eds.), Academic, Orlando, FL, pp. 291–319.
Goodchild, J. (1990) Conjugates of oligonucleotides and modified oligonucleotides: a review of their synthesis and properties. Bioconj. Chem, 1, 165–187.
Chambers, R W. and Moffatt, J. G. (1958) The synthesis of adenosine 5′ and uridine-5′ phosphoramidates. J. Am. Chem. Soc. 80, 3752–3756.
Gilham, P. T. (1968) The synthesis of celluloses containing covalently bound nucleotides, polynucleotides, and nucleic acids. Biochemistry 7, 2809–2813.
Moffatt, J. G. and Khorana, H. G. (1961) Nucleoside polyphosphates. X. The synthesis and some reactions of nucleoside-5′-phosphoromorpholidates and related compounds. Improved methods for the preparation of nucleoside-5′-polyphosphates. J. Am. Chem. Soc. 83, 649–658.
Ivanovskaya, M. G., Gottikh, M. B., and Shabarova, Z. A. (1982) DNA-like duplexes containing repeats. IV. Template-directed polymerization of decade-oxyribonucleotide imidazole. Bioorgan. Khim. 8, 940–944.
Chu, B. C. F., Wahl, G. M., and Orgel, L. E. (1983) Derivatization of unprotected polynucleotides. Nucleic Acids Res. 11, 6513–6529.
Chu, B. C. F. and Orgel, L. E. (1985) Detection of specific DNA sequences with short biotin-labeled probes. DNA 4, 327–331.
Cholet, A. and Kawashima, E. H. (1985) Biotin-labeled synthetic oligodeoxy-ribonucleotides: chemical synthesis and uses as hybridization probes. Nucleic Acids Res. 13, 1529–1541.
Chu, B. C. F. and Orgel, L. E. (1988) Ligation of oligonucleotides to nucleic acids or proteins via disulfide bonds. Nucleic Acids Res. 16, 3671–3691.
Ghosh, S. S., Kao, P. M., and Kwoh, D. Y. (1989) Synthesis of 5′-oligonucle-otide hydrazide derivatives and their use in preparation of enzyme-nucleic acid hybridization probes. Anal. Biochem. 178, 43–51.
Hodges, R. R., Conway, N. E., and McLaughlin, L. W. (1989) “Post-assay” covalent labeling of phosphorothioate-containing nucleic acids with multiple fluorescent markers Biochemistry 28, 261–267
Chu, B. C. F and Orgel, L. E. (1990) A simple procedure for cross-linking complementary oligonucleotides. DNA and Cell Biol. 9, 71–76.
Chu, B. C. F. and Orgel, L. E. (1990) Optimization of the efficiency of cross-linking PtII oligonucleotide phosphorothioate complexes to complementary oligonucleotides. Nucleic Acids Res. 18, 5163–5171
Weith, H. L. and Gilham, P. T (1967) Structural analysis of polynucleotides by sequential base elimination. The sequence of the terminal decanucleotide fragment of the ribonucleic acid from bacteriophage f2. J. Am. Chem. Soc. 89, 5473,5474
Deng, G.-R. and Wu, R. (1983) Terminal transferase: use in the tailing of DNA and for in vitro mutagenesis, in Methods in Enzymology, vol. 100 (Wu, R., Grossman, L., and Moldave, K., eds.). Academic, New York, pp. 96–116.
Stribling, R. (1991) High-performance liquid chromatography of oligogua-nylates at high pH J Chromatog. 538, 474–479.
Gilham, P T. (1962) An addition reaction specific for uridine and guanosine nucleotides and its apphcation to the modification of ribonuclease action J Am Chem. Soc. 84, 687–688
Chu, B. C. F., Kramer, F. R., and Orgel, L. E. (1986) Synthesis of an amplifiable reporter RNA for bioassays. Nucleic Acids Res. 14, 5591–5603
Maniatis, T., Fritsch, E. F., and Sambrook, J. (1982) Molecular Clomng 2nd Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, p. 5,68
Chu, B. C. F. and Orgel, L. E, (1984) Preparation of ligation intermediates and related polynucleotide pyrophosphates. Biochim. Biophys. Acta 782, 103–105.
Joyce, G. F., Inoue, T., and Orgel, L. E. (1984) Non-enzymatic template-directed synthesis on RNA random copolymers poly(C, U) templates. J Mol. Biol. 176, 279–306.
Mukaiyama, T. and Hashimoto, M. (1971) Phosphorylation by oxidation-reduction condensation. Preparation of active phosphorylating reagents. Bull. Chem. Soc. Japan 47, 2284.
Maniatis, T., Fntsch, E. F., and Sambrook, J. (1982) Molecular Cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Chu, B.C. F and Orgel, L. E (1985) Nonenzymatic sequence-specific cleavage of single-stranded DNA Proc. Natl. Acad Sci. USA 82, 963–967
Dreyer, G. B and Dervan, P B. (1985) Sequence-specific cleavage of single-stranded DNA, Oligodeoxynucleotide-EDTA-Fe(II) Proc. Natl. Acad. Sci. USA 82, 968–972.
Chen, C.-H. B. and Sigman, D. S (1986) Nuclease activity of 1,10-phenanthroline-copper: sequence-specific targeting. Proc. Natl. Acad. Sci. USA 83, 7147–7151
LeMaitre, M., Bayard, B, and Lebleu, B. (1987) Specific antiviral activity of a poly(L-Iysine)-conjugated oligodeoxynbonucleotide sequence complementary to vesicular stomatitis virus N protein mRNA initiation site Proc. Natl. Acad Sci. USA 84, 648–652.
Chu, B C. F. and Orgel, L E. (1989) Inhibition of DNA synthesis by cross-linking the template to platinum-thiol derivatives of complementary oligodeoxy-nucleotides. Nucleic Acids Res. 17, 4783–4798.
Teare, J. and Wollenzien, P. (1989) Specificity of site directed psoralen addition to RNA. Nucleic Acids Res. 17, 3359–3372.
King, T. P., Li, Y., and Kochoumian, L. (1978) Preparation of protein conjugates via intermolecular disulfide bond formation. Biochemistry 17, 1499–1506
Blattler, W. A., Kuenzi, B. S., Lambert, J. M., and Senter, P. D. (1985) New heterobifunctional protein cross-linking reagent that forms an acid-labile link. Biochemistry 2A, 1517–1524.
Tsang, V. C. W., Peralta, J. M., and Simons, A. R. (1983) Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis, in Methods in Enzymology, vol. 92 (Langone, J. J. and Van Vunalcis, H., eds.), Academic, New York, pp. 377–391.
EUman, G. I (1959) Tissue sulfhydryl groups. Arch. Biochem. Biophys 82, 70–77.
Murakami, A., Tada, J., Yamagata, K., and Takano, J. (1989) Highly sensitive detection of DNA using enzyme-linked DNA-probe. 1. Colorimetric and fluorometric detection. Nucleic Acids Res. 17, 5587–5595.
Author information
Authors and Affiliations
Rights and permissions
Copyright information
© 1994 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Chu, B.C.E., Orgel, L.E. (1994). Postsynthesis Functionalization of Oligonucleotides. In: Protocols for Oligonucleotide Conjugates. Methods in Molecular Biology, vol 26. Humana Press. https://doi.org/10.1007/978-1-59259-513-6_5
Download citation
DOI: https://doi.org/10.1007/978-1-59259-513-6_5
Publisher Name: Humana Press
Print ISBN: 978-0-89603-252-1
Online ISBN: 978-1-59259-513-6
eBook Packages: Springer Protocols