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A DNA Aptamer for Theophylline with Ultrahigh Selectivity Reminiscent of the Classic RNA Aptamer
| Description: | Since the report of the RNA aptamer for theophylline, theophylline has become a key molecule in chemical biology for designing RNA switches and riboswitches. In addition, theophylline is an important drug for treating airway diseases including asthma. The classic RNA aptamer with excellent selectivity for theophylline has been used to design biosensors, although DNA aptamers are more desirable for stability and cost considerations. In this work, we selected DNA aptamers for theophylline, and all the top sequences shared the same binding motifs. Binding was confirmed using isothermal titration calorimetry and a nuclease digestion assay, showing a dissociation constant (Kd) around 0.5 μM theophylline. The Theo2201 aptamer can be truncated down to 23-mer while still has a Kd of 9.8 μM. The selectivity for theophylline over caffeine is around 250,000-fold based on a strand-displacement assay, which was more than 20-fold higher compared to the classic RNA aptamer. For other tested analogs, the DNA aptamer also showed better selectivity. Using the structure-switching aptamer sensor design method, a detection limit of 17 nM theophylline was achieved in the selection buffer, and a detection limit of 31 nM was obtained in 10% serum. |
| Authors: | Huang, Po-Jung Jimmy; University of Waterloo;  0000-0003-3436-9968Liu, Juewen; University of Waterloo; 0000-0001-5918-9336 |
| Keywords: | DNA SELEX Aptamer Theophylline Biosensor RNA |
| Field of Research: | Biological sciences > Biochemistry > Analytical biochemistry
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| Publication Date: | 2023-04-25 |
| Publisher: | Federated Research Data Repository / dépôt fédéré de données de recherche |
| Funder: | Natural Sciences and Engineering Research Council of Canada Canada First Research Excellence Fund (CFREF); Global Water Futures Program (GWF) |
| URI: | https://doi.org/10.20383/102.0714 |
| Related Identifiers: |
This dataset is cited by
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| Appears in Collections: | Water Institute |
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Creative Commons Public Domain Dedication (CC0 1.0) https://creativecommons.org/publicdomain/zero/1.0/
Citation
Huang, P., Liu, J. (2023). A DNA Aptamer for Theophylline with Ultrahigh Selectivity Reminiscent of the Classic RNA Aptamer. Federated Research Data Repository. https://doi.org/10.20383/102.0714
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