PUB - Publikationen an der Universität Bielefeld

Post-synthetic modification of nucleic acid structures with clickable-functionality is a versatile tool that facilities many emerging applications, including immune evasion, enhancements in stability, fluorescent labelling, chemical 5'-RNA-capping and development of functional aptamers. While certain chemoenzymatic approaches for 3'-azido and alkynyl-labelling are known, equivalent 5'-strategies are either inefficient, complex, or require harsh chemical conditions. Here, we present a modular and facile technology to consecutively modify DNA and RNA strands at both ends with click-modifiable functional groups. Our approach using gamma-modified ATP analogues facilitates T4 PNK catalysed 5'-modification of oligonucleotides, a process which is compatible with TdT-catalysed 3'-elongation using 3'-Azido-2',3'-ddGTP. Finally, we demonstrate that our approach is suitable for both oligo-oligo ligations, as well ssDNA-circularization. We anticipate that such approaches will pave the way for the synthesis of highly functionalized oligonucleotides, improving the therapeutic and diagnostic applicability of oligonucleotides such as in the realm of next-generation-sequencing. © 2023 Wiley-VCH GmbH.