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The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers
Inspired by the chemical synthesis of molecularly imprinted polymers, we demonstrated for the first time, the protein‐target mediated synthesis of enzyme‐generated aptamers (EGAs). We prepared pre‐polymerisation mixtures containing different ratios of nucleotides, an initiator sequence and protein t...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9314878/ https://www.ncbi.nlm.nih.gov/pubmed/35167174 http://dx.doi.org/10.1002/anie.202201061 |
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author | Vacacela, Julio Schaap‐Johansen, Anna‐Lisa Manikova, Patricia Marcatili, Paolo Prado, Marta Sun, Yi Ashley, Jon |
author_facet | Vacacela, Julio Schaap‐Johansen, Anna‐Lisa Manikova, Patricia Marcatili, Paolo Prado, Marta Sun, Yi Ashley, Jon |
author_sort | Vacacela, Julio |
collection | PubMed |
description | Inspired by the chemical synthesis of molecularly imprinted polymers, we demonstrated for the first time, the protein‐target mediated synthesis of enzyme‐generated aptamers (EGAs). We prepared pre‐polymerisation mixtures containing different ratios of nucleotides, an initiator sequence and protein template and incubated each mixture with terminal deoxynucleotidyl transferase (TdT). Upon purification and rebinding of the EGAs against the target, we observed an enhancement in binding of templated‐EGAs towards the target compared to a non‐templated control. These results demonstrate the presence of two primary mechanisms for the formation of EGAs, namely, the binding of random sequences to the target as observed in systematic evolution of ligands by exponential enrichment (SELEX) and the dynamic competition between TdT enzyme and the target protein for binding of EGAs during synthesis. The latter mechanism serves to increase the stringency of EGA‐based screening and represents a new way to develop aptamers that relies on rational design. |
format | Online Article Text |
id | pubmed-9314878 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-93148782022-07-30 The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers Vacacela, Julio Schaap‐Johansen, Anna‐Lisa Manikova, Patricia Marcatili, Paolo Prado, Marta Sun, Yi Ashley, Jon Angew Chem Int Ed Engl Communications Inspired by the chemical synthesis of molecularly imprinted polymers, we demonstrated for the first time, the protein‐target mediated synthesis of enzyme‐generated aptamers (EGAs). We prepared pre‐polymerisation mixtures containing different ratios of nucleotides, an initiator sequence and protein template and incubated each mixture with terminal deoxynucleotidyl transferase (TdT). Upon purification and rebinding of the EGAs against the target, we observed an enhancement in binding of templated‐EGAs towards the target compared to a non‐templated control. These results demonstrate the presence of two primary mechanisms for the formation of EGAs, namely, the binding of random sequences to the target as observed in systematic evolution of ligands by exponential enrichment (SELEX) and the dynamic competition between TdT enzyme and the target protein for binding of EGAs during synthesis. The latter mechanism serves to increase the stringency of EGA‐based screening and represents a new way to develop aptamers that relies on rational design. John Wiley and Sons Inc. 2022-03-04 2022-04-19 /pmc/articles/PMC9314878/ /pubmed/35167174 http://dx.doi.org/10.1002/anie.202201061 Text en © 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes. |
spellingShingle | Communications Vacacela, Julio Schaap‐Johansen, Anna‐Lisa Manikova, Patricia Marcatili, Paolo Prado, Marta Sun, Yi Ashley, Jon The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers |
title | The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers |
title_full | The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers |
title_fullStr | The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers |
title_full_unstemmed | The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers |
title_short | The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers |
title_sort | protein‐templated synthesis of enzyme‐generated aptamers |
topic | Communications |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9314878/ https://www.ncbi.nlm.nih.gov/pubmed/35167174 http://dx.doi.org/10.1002/anie.202201061 |
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