A Kinetic and Fluorogenic Enhancement Strategy for Labeling of Nucleic Acids.
Morten O LoehrNathan W LuedtkePublished in: Angewandte Chemie (International ed. in English) (2022)
Chemical modification of nucleic acids in living cells can be sterically hindered by tight packing of bioorthogonal functional groups in chromatin. To address this limitation, we report here a dual enhancement strategy for nucleic acid-templated reactions utilizing a fluorogenic intercalating agent capable of undergoing inverse electron-demand Diels-Alder (IEDDA) reactions with DNA containing 5-vinyl-2'-deoxyuridine (VdU) or RNA containing 5-vinyl-uridine (VU). Reversible high-affinity intercalation of a novel acridine-tetrazine conjugate "PINK" (K D =5±1 μM) increases the reaction rate of tetrazine-alkene IEDDA on duplex DNA by 60 000-fold (590 M -1 s -1 ) as compared to the non-templated reaction. At the same time, loss of tetrazine-acridine fluorescence quenching renders the reaction highly fluorogenic and detectable under no-wash conditions. This strategy enables live-cell dynamic imaging of acridine-modified nucleic acids in dividing cells.
Keyphrases
- nucleic acid
- single molecule
- living cells
- fluorescent probe
- circulating tumor
- induced apoptosis
- cell free
- high resolution
- gene expression
- electron transfer
- dna damage
- blood brain barrier
- transcription factor
- cell cycle arrest
- oxidative stress
- endoplasmic reticulum stress
- genome wide
- cancer therapy
- cell death
- mass spectrometry
- pi k akt
- fluorescence imaging
- circulating tumor cells