Chemical ligation reactions of oligonucleotides (ONs) have been utilized for various purposes such as non-enzymatic sequences of ONs and intracellular ON-detection etc. Among them, automatic ligation reactions, which preclude the use of activating reagents in the ligation reactions, are useful because of high reaction efficiency and selectivity. For these reactions, phosphorothioate group (PS) has been widely exploited. However, PS group has been used only as a nucleophile, thus limiting the variety of the reaction patterns, resulting linkage structure and its biotechnical application. We herein report a new automatic ligation reaction for oligonucleotides.<br/>The reaction is based on electrophilic phosphorothioester (EPT) group, which was prepared from PS and an electrophilic activator (Scheme). The activated EPT-oligonucleotides are stable for isolation and storage at low temperature, yet retain high reactivity for the reaction with amino-terminated oligonucleotide on template strand to afford the ligated strand with phosphoramidate linkage under physiological conditions. The reaction was successfully applied to both DNA and RNA strands, in either direction from 3’ to 5’ and 5’ to 3 ‘terminals. Since the reaction afforded the native-like linkage structure; phosphoroamidate, the resulting products would have broad ranges of applications.

Chemical ligation reaction for oligonucleotides based on electrophilic phosphorothioester

255th National Meeting (2018)

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