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...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-01-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253116302839 |
| _version_ | 1819242054521192448 |
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| author | David Porciani Giovanni Signore Laura Marchetti Paolo Mereghetti Riccardo Nifosì Fabio Beltram |
| author_facet | David Porciani Giovanni Signore Laura Marchetti Paolo Mereghetti Riccardo Nifosì Fabio Beltram |
| author_sort | David Porciani |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-23T14:33:42Z |
| format | Article |
| id | doaj.art-e8a536d5c7fd40dfb4dc991ee23e9618 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-12-23T14:33:42Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-e8a536d5c7fd40dfb4dc991ee23e96182022-12-21T17:43:26ZengElsevierMolecular Therapy: Nucleic Acids2162-25312014-01-013C10.1038/mtna.2013.71Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin ReceptorDavid Porciani0Giovanni Signore1Laura Marchetti2Paolo Mereghetti3Riccardo Nifosì4Fabio Beltram5NEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Pisa, ItalyCenter for Nanotechnology Innovation@NEST, Istituto Italiano di Tecnologia, Pisa, ItalyNEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Pisa, ItalyCenter for Nanotechnology Innovation@NEST, Istituto Italiano di Tecnologia, Pisa, ItalyNEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Pisa, ItalyNEST, Scuola Normale Superiore and Istituto Nanoscienze-CNR, Pisa, ItalyThanks 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.http://www.sciencedirect.com/science/article/pii/S2162253116302839aptamer foldingaptamer-mediated endocytosisaptamer rational engineeringtransferrin receptor |
| spellingShingle | David Porciani Giovanni Signore Laura Marchetti Paolo Mereghetti Riccardo Nifosì Fabio Beltram Two Interconvertible Folds Modulate the Activity of a DNA Aptamer Against Transferrin Receptor Molecular Therapy: Nucleic Acids aptamer folding aptamer-mediated endocytosis aptamer rational engineering 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 | aptamer folding aptamer-mediated endocytosis aptamer rational engineering transferrin receptor |
| url | http://www.sciencedirect.com/science/article/pii/S2162253116302839 |
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