Novel Pronucleotide Reporters for bioorthogonal Labeling of Nucleic Acids

Abstract

Recent technological advances in fluorescence microscopy provide the opportunity to visualize cellular or viral processes in living systems with high spatiotemporal resolution. Next to these technological advancements, there was significant progress in the field of catalyst-free bioorthogonal reactions. The inverse electron demand Diels-Alder reaction (DAINV) provides exceptional reaction rates between two reactive partners, while being highly specific, thus enabling the tracking of labeled biomolecules in living cells. The attachment of synthetic fluorophores, which meet the demand of modern fluorescence microscopy, to biomolecules of interest, provides an exciting opportunity to study live-cell dynamic processes. A significant advantage of the DAINV is its high adjustability regarding reaction kinetics, biocompatibility, and stability. By identifying the individual most suitable reaction partners, this work aims to develop application-orientated nucleoside and pronucleotide reporters, which enable the visualization of cellular and – especially – viral nucleic acids in living systems. Combining the latest developments in the field of bioorthogonal reactions with our expertise in pronucleotide systems achieved an eagerly awaited breakthrough in live-cell imaging of in vivo labeled nucleic acids.