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Versatile synthesis of oligodeoxyribonucleotide–oligospermine conjugates

Nature Protocols volume 2, pages 1360–1367 (2007)Cite this article

Abstract

A protocol for the rapid, automated synthesis of oligospermine–oligonucleotide sequences is described. To this end, a protected spermine phosphoramidite derivative was synthesized in six steps from spermine and used as the fifth synthon in an oligonucleotide synthesizer. Parameters were optimized to reach greater than 95% coupling yields. Cationic oligonucleotides show enhanced hybridization and strand invasion properties, and hence are an alternative to conventional oligonucleotides for molecular biology, diagnostic and potential therapeutic applications. A multi-gram-scale synthesis of the spermine phosphoramidite allowing several hundred coupling steps takes 2–3 weeks. Oligonucleotide synthesis and purification takes approximately 3 d.

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Figure 1: Synthesis of spermine synthon 1 and of oligonucleotide–spermine conjugates.
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Figure 4: Matrix-assisted laser desorption/ionization (MALDI) spectrum (matrix: hydroxypicolinic acid).
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Figure 6
Figure 7: Matrix-assisted laser desorption/ionization (MALDI) spectrum (matrix: hydroxypicolinic acid).

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Authors and Affiliations

  1. Polyplus-transfection SA, Bioparc, Boulevard S. Brandt, BP90018, Illkirch, 67401, France

    Emilie Voirin

  2. Laboratoire de Chimie Génétique associé au C.N.R.S., Université Louis Pasteur de Strasbourg, Faculté de Pharmacie, B.P. 24, Illkirch, 67401, France

    Jean-Paul Behr & Mitsuharu Kotera

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  1. Emilie Voirin
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Correspondence to Mitsuharu Kotera.

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Dr Behr is a minor shareholder of a company that licensed this technology.

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Voirin, E., Behr, JP. & Kotera, M. Versatile synthesis of oligodeoxyribonucleotide–oligospermine conjugates. Nat Protoc 2, 1360–1367 (2007). https://doi.org/10.1038/nprot.2007.177

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