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Chemo‐Enzymatic Synthesis of Position‐Specifically Modified RNA for Biophysical Studies including Light Control and NMR Spectroscopy
Author(s) -
Keyhani Sara,
Goldau Thomas,
Blümler Anja,
Heckel Alexander,
Schwalbe Harald
Publication year - 2018
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201807125
Subject(s) - nucleotide , rna , oligonucleotide , chemistry , nuclear magnetic resonance spectroscopy , enzyme , riboswitch , stereochemistry , combinatorial chemistry , biochemistry , non coding rna , dna , gene
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.