Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 57, Issue 37, Pages 12017-12021Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201807125
Keywords
chemo-enzymatic; light-control; NMR spectroscopy; photocaged RNA; RNA
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Funding
- Deutsche Forschungsgemeinschaft (DFG) [SFB 902, GRK 1986]
- Cluster of Excellence Macromolecular Complexes [EXC 115]
- state of Hessen
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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 C-13,N-15-labelled nucleosides. All four ribonucleotides containing ortho-nitrophenylethyl (NPE) photocages, photoswitchable azobenzene C-nucleotides and C-13,N-15-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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