Abstract
The aim of this study is to develop a selective adenosine aptamer sensor using a rational approach. Unlike traditional RNA aptamers developed from SELEX, duplex DNA containing an abasic site can function as a general scaffold to rationally design aptamers for small aromatic molecules. We discovered that abasic site-containing triplex DNA can also function as an aptamer and provide better affinity than duplex DNA aptamers. A novel adenosine aptamer sensor was designed using such a triplex. The aptamer is modified with furano-dU in the binding site to sense the binding. The sensor bound adenosine has a dissociation constant of 400 nM, more than tenfold stronger than the adenosine aptamer developed from SELEX. The binding quenched furano-dU fluorescence by 40%. It was also demonstrated in this study that this sensor is selective for adenosine over uridine, cytidine, guanosine, ATP, and AMP. The detection limit of this sensor is about 50 nM. The sensor can be used to quantify adenosine concentrations between 50 nM and 2 μM.
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Acknowledgments
This research was supported by the faculty start-up fund of New Jersey Institute of Technology. We also thank Dr. Robert Donnelly of the University of Medicine and Dentistry of New Jersey for assistance in oligonucleotide synthesis.
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Patel, M., Dutta, A. & Huang, H. A selective adenosine sensor derived from a triplex DNA aptamer. Anal Bioanal Chem 400, 3035–3040 (2011). https://doi.org/10.1007/s00216-011-4996-1
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DOI: https://doi.org/10.1007/s00216-011-4996-1