Design and synthesis of double‐stranded oligonucleotides containing reactive acylphosphate internucleotide groups

FEBS Letters

Kanevsky, I.E¹  Kuznetsova, S.A¹  Shabarova, Z.A¹ 

[1] Chemistry Department, Moscow State University, Moscow 119899, Russia

关键词: Reactive oligonucleotide synthesis;    DNA duplex;    DNA dumbbell;    Template-induced chemical ligation;    Sugar phosphate backbone;    Acylphosphate internucleotide group;    EDC;    1-ethyl-3-(3′-dimethylaminopropyl)carbodiimide;    MeIm;    N-methylimidazole;    EDA;    ethylenediamine;    SPI groups;    substituted pyrophosphate internucleotide groups;    ds;    double-stranded;   

DOI  :  10.1016/S0014-5793(98)00812-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

DNA duplex and dumbbells containing chemically active acylphosphate internucleotide groups were synthesized. To obtain these compounds the chemical ligation method was used. The acylphosphate group was inserted into a DNA duplex and dumbbells as a result of template-directed condensation of 5′-phosphate and especially introduced 3′-carboxy groups of oligonucleotides. 1-Ethyl-3-(3′-dimethylaminopropyl)carbodiimide (EDC) was used as a condensing agent. Oligonucleotides containing a carboxy group were obtained by the interaction of their 3′-phosphate with glycine methyl ester under the action of EDC, followed by ester hydrolysis. The yields of acylphosphate-containing double-stranded oligonucleotides achieved 15–25% depending on the structure of their precursors. It was shown that these compounds are acylating agents and are efficiently cleaved in near-physiological conditions under the action of ethylenediamine or N-methylimidazole. These results indicate that double-stranded oligonucleotides carrying acylphosphate internucleotide groups could constitute new crosslinking reagents for affinity modification of DNA recognizing proteins.

【 授权许可】

Unknown