Solid-Phase-Supported Chemoenzymatic Synthesis of a Light-Activatable tRNA Derivative

Anja Blümler, Harald Schwalbe, Alexander Heckel

Angew. Chem. Int. Ed.
https://doi.org/10.1002/anie.202111613

Here, we present a multi-cycle chemoenzymatic synthesis of modified RNA with simplified solid-phase handling to overcome size limitations of RNA synthesis. It combines the advantages of classical chemical solid-phase synthesis and enzymatic synthesis methods using magnetic streptavidin beads and biotinylated RNA. Successful introduction of light-controllable RNA nucleotides into the tRNAMet sequence was confirmed by gel electrophoresis and mass spectrometry. The methods tolerate modifications in the RNA phosphodiester backbone and allow introductions of photocaged and photoswitchable nucleotides as well as photocleavable strand breaks and fluorophores.