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Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly
Joining oligonucleotides together (ligation) is a powerful means of retrieving information from the nanoscale. To recover this information, the linkages created must be compatible with polymerases. However, enzymatic ligation is restrictive and current chemical ligation methods lack flexibility. Her...
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/PMC7383975/ https://www.ncbi.nlm.nih.gov/pubmed/32153132 http://dx.doi.org/10.1002/anie.202000209 |
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author | Shivalingam, Arun Taemaitree, Lapatrada El‐Sagheer, Afaf H. Brown, Tom |
author_facet | Shivalingam, Arun Taemaitree, Lapatrada El‐Sagheer, Afaf H. Brown, Tom |
author_sort | Shivalingam, Arun |
collection | PubMed |
description | Joining oligonucleotides together (ligation) is a powerful means of retrieving information from the nanoscale. To recover this information, the linkages created must be compatible with polymerases. However, enzymatic ligation is restrictive and current chemical ligation methods lack flexibility. Herein, a versatile ligation platform based on the formation of urea and squaramide artificial backbones from minimally modified 3′‐ and 5′‐amino oligonucleotides is described. One‐pot ligation gives a urea linkage with excellent read‐through speed, or a squaramide linkage that is read‐through under selective conditions. The squaramide linkage can be broken and reformed on demand, while stable pre‐activated precursor oligonucleotides expand the scope of the ligation reaction to reagent‐free, mild conditions. The utility of our system is demonstrated by replacing the enzymatically biased RNA‐to‐DNA reverse transcription step of RT‐qPCR with a rapid nucleic‐acid‐template‐dependent DNA chemical ligation system, that allows direct RNA detection. |
format | Online Article Text |
id | pubmed-7383975 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-73839752020-07-28 Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly Shivalingam, Arun Taemaitree, Lapatrada El‐Sagheer, Afaf H. Brown, Tom Angew Chem Int Ed Engl Research Articles Joining oligonucleotides together (ligation) is a powerful means of retrieving information from the nanoscale. To recover this information, the linkages created must be compatible with polymerases. However, enzymatic ligation is restrictive and current chemical ligation methods lack flexibility. Herein, a versatile ligation platform based on the formation of urea and squaramide artificial backbones from minimally modified 3′‐ and 5′‐amino oligonucleotides is described. One‐pot ligation gives a urea linkage with excellent read‐through speed, or a squaramide linkage that is read‐through under selective conditions. The squaramide linkage can be broken and reformed on demand, while stable pre‐activated precursor oligonucleotides expand the scope of the ligation reaction to reagent‐free, mild conditions. The utility of our system is demonstrated by replacing the enzymatically biased RNA‐to‐DNA reverse transcription step of RT‐qPCR with a rapid nucleic‐acid‐template‐dependent DNA chemical ligation system, that allows direct RNA detection. John Wiley and Sons Inc. 2020-05-07 2020-07-06 /pmc/articles/PMC7383975/ /pubmed/32153132 http://dx.doi.org/10.1002/anie.202000209 Text en © 2020 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 | Research Articles Shivalingam, Arun Taemaitree, Lapatrada El‐Sagheer, Afaf H. Brown, Tom Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly |
title | Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly |
title_full | Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly |
title_fullStr | Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly |
title_full_unstemmed | Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly |
title_short | Squaramides and Ureas: A Flexible Approach to Polymerase‐Compatible Nucleic Acid Assembly |
title_sort | squaramides and ureas: a flexible approach to polymerase‐compatible nucleic acid assembly |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7383975/ https://www.ncbi.nlm.nih.gov/pubmed/32153132 http://dx.doi.org/10.1002/anie.202000209 |
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