Expanding fluorescent base analogue labelling of long RNA by in vitro transcription.

ABSTRACT: To track RNA inside living cells and to study its natural interactions and mechanisms is vital in RNA research and the development of RNA-based therapeutics. In this work, we show that the fluorescent adenine analogue 2CNqA-triphosphate (2CNqATP) is accepted by the polymerase of a standard in vitro transcription, which results in fluorescently labelled long RNA. We demonstrate that the labelling degree can be tuned by the fractional exchange of natural ATP for 2CNqATP in the in vitro transcription reaction. Furthermore, in a thorough spectroscopic characterization, we present how the fluorescence lifetime and quantum yield change upon incorporation at different labelling degrees and how this in turn affects overall brightness per RNA strand. After transfecting human cells with the correspo

SUBMITTER: Pfeiffer P 

PROVIDER: S-EPMC12664369 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature