Voirin Emilie, Behr Jean-Paul, Kotera Mitsuharu
Polyplus-transfection SA, Bioparc, Boulevard S. Brandt, BP90018, 67401 Illkirch, France.
Nat Protoc. 2007;2(6):1360-7. doi: 10.1038/nprot.2007.177.
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.
本文描述了一种快速、自动合成低聚精胺-寡核苷酸序列的方案。为此，从精胺出发经六步合成了一种受保护的精胺亚磷酰胺衍生物，并将其用作寡核苷酸合成仪中的第五个合成子。对参数进行了优化，以实现大于95%的偶联产率。阳离子寡核苷酸表现出增强的杂交和链入侵特性，因此是用于分子生物学、诊断和潜在治疗应用的传统寡核苷酸的替代品。多克规模合成精胺亚磷酰胺，允许进行数百次偶联步骤，耗时2至3周。寡核苷酸的合成和纯化大约需要3天。
