Chemo-Enzymatic Synthesis of Position-Specifically Modified RNA for Biophysical Studies including Light Control and NMR Spectroscopy

Angewandte Chemie
Sara KeyhaniHarald Schwalbe

Abstract

The investigation of non-coding RNAs requires RNAs containing modifications at every possible position within the oligonucleotide. Here, we present the chemo-enzymatic RNA synthesis containing photoactivatable or 13 C,15 N-labelled nucleosides. All four ribonucleotides containing ortho-nitrophenylethyl (NPE) photocages, photoswitchable azobenzene C-nucleotides and 13 C,15 N-labelled nucleotides were incorporated position-specifically in high yields. We applied this approach for the synthesis of light-inducible 2'dG-sensing riboswitch variants and detected ligand-induced structural reorganization upon irradiation by NMR spectroscopy. This chemo-enzymatic method opens the possibility to incorporate a wide range of modifications at any desired position of RNAs of any lengths beyond the limits of solid-phase synthesis.

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Citations

Mar 16, 2019·RNA Biology·Gerd HanspachMartin Hengesbach
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Jun 12, 2021·Chemistry : a European Journal·Anaïs DepaixJoanna Kowalska
Nov 17, 2019·Journal of the American Chemical Society·Yusuke TakezawaMitsuhiko Shionoya
Nov 6, 2021·Angewandte Chemie·Anja BlümlerAlexander Heckel

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