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Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor
Thanks to their ability to recognize biomolecular targets with high affinity and specificity, nucleic acid aptamers are increasingly investigated as diagnostic and therapeutic tools, particularly when their targets are cell-surface receptors. Here, we investigate the relationship between the folding...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3912326/ https://www.ncbi.nlm.nih.gov/pubmed/24472870 http://dx.doi.org/10.1038/mtna.2013.71 |
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author | Porciani, David Signore, Giovanni Marchetti, Laura Mereghetti, Paolo Nifosì, Riccardo Beltram, Fabio |
author_facet | Porciani, David Signore, Giovanni Marchetti, Laura Mereghetti, Paolo Nifosì, Riccardo Beltram, Fabio |
author_sort | Porciani, David |
collection | PubMed |
description | Thanks to their ability to recognize biomolecular targets with high affinity and specificity, nucleic acid aptamers are increasingly investigated as diagnostic and therapeutic tools, particularly when their targets are cell-surface receptors. Here, we investigate the relationship between the folding of an anti-mouse transferrin receptor DNA aptamer and its interaction with the transferrin receptor both in vitro and in living cells. We identified and purified two aptamer conformers by means of chromatographic techniques. Fluorescence-anisotropy measurements showed that only one fold is able to bind mouse transferrin receptor. Besides displaying enhanced endocytosis in living mouse fibroblasts, the purified active fold is internalized also in human pancreatic cancer cells. Starting from these observations, we rationally designed variations of the parent sequence aimed at stabilizing the active fold, and consequently increase aptamer activity. A truncated version and full-length mutants with higher affinity than the parent sequence are shown. |
format | Online Article Text |
id | pubmed-3912326 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-39123262014-02-04 Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor Porciani, David Signore, Giovanni Marchetti, Laura Mereghetti, Paolo Nifosì, Riccardo Beltram, Fabio Mol Ther Nucleic Acids Original Article Thanks to their ability to recognize biomolecular targets with high affinity and specificity, nucleic acid aptamers are increasingly investigated as diagnostic and therapeutic tools, particularly when their targets are cell-surface receptors. Here, we investigate the relationship between the folding of an anti-mouse transferrin receptor DNA aptamer and its interaction with the transferrin receptor both in vitro and in living cells. We identified and purified two aptamer conformers by means of chromatographic techniques. Fluorescence-anisotropy measurements showed that only one fold is able to bind mouse transferrin receptor. Besides displaying enhanced endocytosis in living mouse fibroblasts, the purified active fold is internalized also in human pancreatic cancer cells. Starting from these observations, we rationally designed variations of the parent sequence aimed at stabilizing the active fold, and consequently increase aptamer activity. A truncated version and full-length mutants with higher affinity than the parent sequence are shown. Nature Publishing Group 2014-01 2014-01-28 /pmc/articles/PMC3912326/ /pubmed/24472870 http://dx.doi.org/10.1038/mtna.2013.71 Text en Copyright © 2014 The American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-sa/3.0/ Molecular Therapy-Nucleic Acids is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/ |
spellingShingle | Original Article Porciani, David Signore, Giovanni Marchetti, Laura Mereghetti, Paolo Nifosì, Riccardo Beltram, Fabio Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor |
title | Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor |
title_full | Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor |
title_fullStr | Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor |
title_full_unstemmed | Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor |
title_short | Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor |
title_sort | two interconvertible folds modulate the activity of a dna aptamer against transferrin receptor |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3912326/ https://www.ncbi.nlm.nih.gov/pubmed/24472870 http://dx.doi.org/10.1038/mtna.2013.71 |
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