General Method for Post-Synthetic Modification of Oligonucleotides Based on Oxidative Amination of 4-Thio-2′-deoxyuridine

Functionalized oligonucleotides (ONs) are widely applied as target binding molecules for biosensing and regulators for gene expression. Numerous efforts have been focused on developing facile methods for preparing these useful ONs carrying diverse modifications. Herein, we present a general method for postsynthetic modification of ONs via oxidative amination of 4-thio-2'-deoxyuridine (4SdU). 4SdU-containing ON can be derived by both alkyl and aromatic amines. Using this approach, ONs are successfully attached with alkyne/azide, biotin and dansylamide moieties, and these as-prepared ONs possess the expected biorthogonal reactivity, streptavidin affinity and fluorescent property, respectively. Furthermore, we also directly install fluorophores to the ON nucleobase based on oxidative amination of 4SdU, and these fluorophores exhibit distinct luminescence behaviors before and after conjugation. We believe our method will be a versatile strategy for constructing various functionalized ONs used in a wide range of nucleic acid applications.

Automated summary

A general method for postsynthetic modification of ONs via oxidative amination of 4-thio-2'-deoxyuridine (4SdU) is presented, allowing successful attachment of alkyne/azide, biotin, and dansylamide moieties to ONs. Fluorophores can also be directly installed to the ON nucleobase based on this method. The versatile strategy is believed to be useful for constructing various functionalized ONs in nucleic acid applications.