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A highly specific sodium aptamer probed by 2-aminopurine for robust Na+ sensing

dc.contributor.authorZhou, Wenhu
dc.contributor.authorDing, Jinsong
dc.contributor.authorLiu, Juewen
dc.date.accessioned2017-04-28T16:12:00Z
dc.date.available2017-04-28T16:12:00Z
dc.date.issued2016-09-20
dc.description.abstractSodium is one of the most abundant metals in the environment and in biology, playing critical ecological and physiological roles. Na+ is also the most common buffer salt for nucleic acids research, while its specific interaction with DNA has yet to be fully studied. Herein, we probe a highly selective and robust Na+ aptamer using 2-aminopurine (2AP), a fluorescent adenine analog. This aptamer has two DNA strands derived from the Ce13d DNAzyme. By introducing a 2AP at the cleavage site of the substrate strand, Na+ induces similar to 40% fluorescence increase. The signaling is improved by a series of rational mutations, reaching > 600% with the C(10)A(20) double mutant. This fluorescence enhancement suggests relaxed base stacking near the 2AP label upon Na+ binding. By replacing a non-conserved adenine in the enzyme strand by 2AP, Na+-dependent fluorescence quenching is observed, suggesting that the enzyme loop folds into a more compact structure upon Na+ binding. The fluorescence changes allow for Na+ detection. With an optimized sequence, a detection limit of 0.4 mM Na+ is achieved, reaching saturated signal in less than 10 s. The sensor response is insensitive to ionic strength, which is critical for Na+ detection.en
dc.description.sponsorshipUniversity of Waterloo, the Natural Sciences and Engineering Research Council of Canada (NSERC); Foundation for Shenghua Scholar of Central South University and the National Natural Science Foundation of China [21301195]; Fellowship from the China Scholarship Council (CSC) [201406370116 to W.Z.]. Funding for open access charge: Natural Sciences and Engineering Research Council of Canada (NSERC).en
dc.identifier.urihttp://dx.doi.org/10.1093/nar/gkw845
dc.identifier.urihttp://hdl.handle.net/10012/11798
dc.language.isoenen
dc.publisherOxford University Pressen
dc.rightsAttribution-NonCommercial 4.0 International*
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/*
dc.subjectLanthanide-Dependent Dnazymeen
dc.subjectIn-Vitro Selectionen
dc.subjectResonance Energy-Transferen
dc.subjectUranyl-Specific Dnazymeen
dc.subjectRNA-Cleaving Dnazymeen
dc.subjectMetal-Ionsen
dc.subjectDna Enzymeen
dc.subjectLiving Cellsen
dc.subjectSitesen
dc.subjectSensorsen
dc.titleA highly specific sodium aptamer probed by 2-aminopurine for robust Na+ sensingen
dc.typeArticleen
dcterms.bibliographicCitationZhou, W., Ding, J., & Liu, J. (2016). A highly specific sodium aptamer probed by 2-aminopurine for robust Na+ sensing. Nucleic Acids Research, 44(21), 10377–10385. https://doi.org/10.1093/nar/gkw845en
uws.contributor.affiliation1Faculty of Scienceen
uws.contributor.affiliation2Chemistryen
uws.contributor.affiliation3Waterloo Institute for Nanotechnology (WIN)en
uws.peerReviewStatusRevieweden
uws.scholarLevelFacultyen
uws.typeOfResourceTexten

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