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Genetic Code Expansion Facilitates Position-Selective Labeling of RNA for Biophysical Studies.

Authors :
Hegelein A
Müller D
Größl S
Göbel M
Hengesbach M
Schwalbe H
Source :
Chemistry (Weinheim an der Bergstrasse, Germany) [Chemistry] 2020 Feb 06; Vol. 26 (8), pp. 1800-1810. Date of Electronic Publication: 2020 Jan 21.
Publication Year :
2020

Abstract

Nature relies on reading and synthesizing the genetic code with high fidelity. Nucleic acid building blocks that are orthogonal to the canonical A-T and G-C base-pairs are therefore uniquely suitable to facilitate position-specific labeling of nucleic acids. Here, we employ the orthogonal kappa-xanthosine-base-pair for in vitro transcription of labeled RNA. We devised an improved synthetic route to obtain the phosphoramidite of the deoxy-version of the kappa nucleoside in solid phase synthesis. From this DNA template, we demonstrate the reliable incorporation of xanthosine during in vitro transcription. Using NMR spectroscopy, we show that xanthosine introduces only minor structural changes in an RNA helix. We furthermore synthesized a clickable 7-deaza-xanthosine, which allows to site-specifically modify transcribed RNA molecules with fluorophores or other labels.<br /> (© 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

Details

Language :
English
ISSN :
1521-3765
Volume :
26
Issue :
8
Database :
MEDLINE
Journal :
Chemistry (Weinheim an der Bergstrasse, Germany)
Publication Type :
Academic Journal
Accession number :
31692134
Full Text :
https://doi.org/10.1002/chem.201904623