Smalltalks "How to make nucleic acids shine"

Abstract:

Remember COVID-19 and how it led to the first RNA-vaccine coming into play?
The field of ribonucleic acid (RNA)-based therapeutics is evolving rapidly and the development of more accurate tools to study them and their true action in cells is vital. In my work, I investigate fluorescent nucleobase analogues (FBAs) as building blocks to make RNA detectable by fluorescence-based readout techniques, like fluorescence microscopy. Our FBAs offer unique possibilities to probe nucleic acids, like RNA, at physiological conditions while maintaining their natural interaction patterns. They are accepted as substrates for enzymatic reactions and hence can be used as internal labels of functional nucleic acids. We also present the spontaneous cellular uptake of two FBAs as nucleoside triphosphates. Relatively small differences in the molecular structure of the modified nucleobases result in significant differences in their intracellular localization. Following the accumulation in living cells we have found one base analogue to be accepted as a building block and incorporated into RNA, rendering the cellular RNA fluorescent. The spontaneous uptake into living cells and RNA incorporation of FBA triphosphates along with in vitro labeling of long RNAs, and the ability of FBAs to maintain native nucleic acid properties, offer a versatile toolbox to study the dynamic aspects of RNA-based drug delivery and trafficking within living cells.