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Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies
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‐xant...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7027469/ https://www.ncbi.nlm.nih.gov/pubmed/31692134 http://dx.doi.org/10.1002/chem.201904623 |
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author | Hegelein, Andreas Müller, Diana Größl, Sylvester Göbel, Michael Hengesbach, Martin Schwalbe, Harald |
author_facet | Hegelein, Andreas Müller, Diana Größl, Sylvester Göbel, Michael Hengesbach, Martin Schwalbe, Harald |
author_sort | Hegelein, Andreas |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-7027469 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-70274692020-02-24 Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies Hegelein, Andreas Müller, Diana Größl, Sylvester Göbel, Michael Hengesbach, Martin Schwalbe, Harald Chemistry Full Papers 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. John Wiley and Sons Inc. 2020-01-21 2020-02-06 /pmc/articles/PMC7027469/ /pubmed/31692134 http://dx.doi.org/10.1002/chem.201904623 Text en © 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Full Papers Hegelein, Andreas Müller, Diana Größl, Sylvester Göbel, Michael Hengesbach, Martin Schwalbe, Harald Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies |
title | Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies |
title_full | Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies |
title_fullStr | Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies |
title_full_unstemmed | Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies |
title_short | Genetic Code Expansion Facilitates Position‐Selective Labeling of RNA for Biophysical Studies |
title_sort | genetic code expansion facilitates position‐selective labeling of rna for biophysical studies |
topic | Full Papers |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7027469/ https://www.ncbi.nlm.nih.gov/pubmed/31692134 http://dx.doi.org/10.1002/chem.201904623 |
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