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However, this covalent labeling approach suffers from several limitations, including complex and costly chemical modification, incompatibility with long aptamer sequences, and susceptibility to false-positive signals. To address these limitations, we constructed a novel aptamer-based fluorescent sensor using Lettuce indicator, a light-up DNA aptamer that adopts a precise three-dimensional structure, which enables it to selectively bind and activate the otherwise non-fluorescent small-molecule fluorophores. In our design, we destabilized Lettuce by fusing it with a target-binding aptamer via a transducer sequence. Upon target binding, structural rearrangement is triggered through the transducer, leading to the folding of Lettuce and restoration of its ability to activate the fluorophore, generating a target-dependent fluorescent signal. Through systematic optimization of the transducer and target-binding aptamer sequences, we created sensors for diverse targets, including small molecules, proteins, and metal ions. These sensors exhibit high signal-to-noise ratios, sensitivity and selectivity, and a wide dynamic range. As a proof-of-concept demonstration, a paper-based test strip for cost-effective and rapid detection of small molecule mycotoxins was developed. This versatile design provides a generalizable platform for the development of aptamer-based sensors, opening the way for future detection of diverse analytes.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c03182","type":"journal-article","created":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T16:01:56Z","timestamp":1789228916000},"page":"28158-28170","source":"Crossref","is-referenced-by-count":0,"title":["Aptamer-Based Sensing with Fluorescent Light-Up DNA"],"prefix":"10.1021","volume":"98","author":[{"given":"Xin","family":"Ji","sequence":"first","affiliation":[{"name":"Beijing Technology and Business University , , No. 11 Fucheng Road , Haidian District, ,","place":["Beijing, China, 100048"]},{"name":"Institute of Zoology, Chinese Academy of Sciences , , No. 1 West Beichen Road , Chaoyang District, ,","place":["Beijing, China, 100101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ranran","family":"Hou","sequence":"additional","affiliation":[{"name":"Beijing Technology and Business University , , No. 11 Fucheng Road , Haidian District, ,","place":["Beijing, China, 100048"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hai","family":"Bao","sequence":"additional","affiliation":[{"name":"Beijing Technology and Business University , , No. 11 Fucheng Road , Haidian District, ,","place":["Beijing, China, 100048"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Liu","sequence":"additional","affiliation":[{"name":"Institute of Zoology, Chinese Academy of Sciences , , No. 1 West Beichen Road , Chaoyang District, ,","place":["Beijing, China, 100101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuoxin","family":"Yao","sequence":"additional","affiliation":[{"name":"Hunan Normal University , , No. 36 Lushan Road , Yuelu District, ,","place":["Changsha, China, 410081"]},{"name":"Institute of Subtropical Agriculture, Chinese Academy of Sciences , ,","place":["Changsha, China, 410125"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chi","family":"Chen","sequence":"additional","affiliation":[{"name":"Beijing Technology and Business University , , No. 11 Fucheng Road , Haidian District, ,","place":["Beijing, China, 100048"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yulong","family":"Yin","sequence":"additional","affiliation":[{"name":"Institute of Subtropical Agriculture, Chinese Academy of Sciences , ,","place":["Changsha, China, 410125"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Liuqin","family":"He","sequence":"additional","affiliation":[{"name":"Hunan Normal University , , No. 36 Lushan Road , Yuelu District, ,","place":["Changsha, China, 410081"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3149-9783","authenticated-orcid":true,"given":"Xing","family":"Li","sequence":"additional","affiliation":[{"name":"Institute of Zoology, Chinese Academy of Sciences , , No. 1 West Beichen Road , Chaoyang District, ,","place":["Beijing, China, 100101"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3709-3143","authenticated-orcid":true,"given":"Xiaoying","family":"Zhao","sequence":"additional","affiliation":[{"name":"Beijing Technology and Business University , , No. 11 Fucheng Road , Haidian District, ,","place":["Beijing, China, 100048"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,12]]},"reference":[{"key":"2026092909094332600_cit1","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1146\/annurev-pharmtox-010716-104558","article-title":"Aptamers\nas Therapeutics","volume":"57","author":"Nimjee","year":"2017","journal-title":"Annu. Rev. Pharmacol. Toxicol."},{"issue":"2","key":"2026092909094332600_cit2","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1016\/j.trac.2007.12.004","article-title":"Aptamer-based\nbiosensors","volume":"27","author":"Song","year":"2008","journal-title":"TrAC, Trends Anal. Chem."},{"issue":"10","key":"2026092909094332600_cit3","doi-asserted-by":"publisher","first-page":"12886","DOI":"10.1021\/acsomega.8b01812","article-title":"Label-free\naptamer-based biosensor for specific detection of chloramphenicol\nusing AIE probe and graphene oxide","volume":"3","author":"Zhang","year":"2018","journal-title":"Acs Omega"},{"issue":"6","key":"2026092909094332600_cit4","doi-asserted-by":"publisher","DOI":"10.3390\/bios12060423","article-title":"Development of Fluorescent Aptasensors Based on G-Quadruplex Quenching\nAbility for Ochratoxin A and Potassium Ions Detection","volume":"12","author":"Yang","year":"2022","journal-title":"Biosensors"},{"issue":"5","key":"2026092909094332600_cit5","doi-asserted-by":"publisher","first-page":"4541","DOI":"10.3390\/s100504541","article-title":"Design strategies for aptamer-based\nbiosensors","volume":"10","author":"Han","year":"2010","journal-title":"Sensors"},{"issue":"2","key":"2026092909094332600_cit6","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1021\/ac1025896","article-title":"Simple\nand convenient G-quadruplex-based turn-on fluorescence assay for 3\u2032\u2192\n5\u2032 exonuclease activity","volume":"83","author":"Leung","year":"2011","journal-title":"Anal. 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Biol."},{"issue":"1","key":"2026092909094332600_cit19","doi-asserted-by":"publisher","first-page":"246","DOI":"10.1021\/acssensors.4c02331","article-title":"Design of Label-Free\nDNA Light-Up Aptaswitches for\nMultiplexed Biosensing","volume":"10","author":"Osman","year":"2025","journal-title":"ACS Sens."},{"issue":"14","key":"2026092909094332600_cit20","doi-asserted-by":"publisher","first-page":"7666","DOI":"10.1093\/nar\/gkad541","article-title":"Structural\nbasis for\nhigh-affinity recognition of aflatoxin B1 by a DNA aptamer","volume":"51","author":"Xu","year":"2023","journal-title":"Nucleic Acids Res."},{"issue":"3","key":"2026092909094332600_cit21","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1002\/bab.1586","article-title":"Prediction of shorter\noligonucleotide sequences recognizing aflatoxin M1","volume":"65","author":"Pandey","year":"2018","journal-title":"Biotechnol. Appl. 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Bioelectron."},{"key":"2026092909094332600_cit27","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1002\/9781118971147.ch1","article-title":"ICHQ1A(R2) Stability\nTesting of New Drug Substance and Product and ICHQ1C Stability Testing\nof New Dosage Forms","volume-title":"ICH Quality Guidelines:\nAn Implementation Guide","author":"Rignall","year":"2017"},{"issue":"6073","key":"2026092909094332600_cit28","doi-asserted-by":"publisher","first-page":"1194","DOI":"10.1126\/science.1218298","article-title":"Fluorescence imaging\nof cellular metabolites with RNA","volume":"335","author":"Paige","year":"2012","journal-title":"Science"},{"key":"2026092909094332600_cit29a","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2020.00676","article-title":"Time-resolved fluorescence\nimmunochromatography assay (TRFICA) for aflatoxin: aiming at increasing\nstrip method sensitivity","volume":"11","author":"Li","year":"2020","journal-title":"Fronti. 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Chem."},{"key":"2026092909094332600_cit31","doi-asserted-by":"publisher","DOI":"10.3389\/fchem.2021.653869","article-title":"Aptamer-Based\nFluorescence\nQuenching Approach for Detection of Aflatoxin M(1) in Milk","volume":"9","author":"Qiao","year":"2021","journal-title":"Front. Chem."},{"issue":"24","key":"2026092909094332600_cit32","doi-asserted-by":"publisher","first-page":"6269","DOI":"10.1007\/s00216-018-1237-x","article-title":"Development of aptamer fluorescent\nswitch assay for\naflatoxin B1 by using fluorescein-labeled aptamer and black hole quencher\n1-labeled complementary DNA","volume":"410","author":"Li","year":"2018","journal-title":"Anal. Bioanal.\nChem."},{"key":"2026092909094332600_cit33","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1016\/j.foodcont.2014.12.008","article-title":"Detection\nof aflatoxin B1 by aptamer-based\nbiosensor using PAMAM dendrimers as immobilization platform","volume":"52","author":"Castillo","year":"2015","journal-title":"Food Control"},{"issue":"5","key":"2026092909094332600_cit34","doi-asserted-by":"publisher","first-page":"2724","DOI":"10.1016\/j.bios.2010.09.032","article-title":"Aptamer-based colorimetric\nbiosensing of Ochratoxin A using unmodified gold nanoparticles indicator","volume":"26","author":"Yang","year":"2011","journal-title":"Biosens. Bioelectron."},{"issue":"4","key":"2026092909094332600_cit35","doi-asserted-by":"publisher","DOI":"10.1007\/s00604-019-3325-9","article-title":"A rapid fluorometric method for determination\nof aflatoxin B1 in\nplant-derived food by using a thioflavin T-based aptasensor","volume":"186","author":"Li","year":"2019","journal-title":"Microchim. Acta"},{"key":"2026092909094332600_cit36","doi-asserted-by":"publisher","DOI":"10.1016\/j.dyepig.2019.107788","article-title":"Label-free G-Quadruplex aptamer and\nThioflavin-T based turn-off fluorescent\ndetection of ethanolamine","volume":"172","author":"Bayrac\u0327","year":"2020","journal-title":"Dyes Pigm."},{"key":"2026092909094332600_cit37","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2020.00080","article-title":"Development of ELISA and lateral\nflow immunoassays for ochratoxins\n(OTA and OTB) detection based on monoclonal antibody","volume":"10","author":"Fadlalla","year":"2020","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"2026092909094332600_cit38","doi-asserted-by":"publisher","DOI":"10.1016\/j.foodchem.2019.125851","article-title":"Aptamer assisted ultrafiltration\ncleanup with high\nperformance liquid chromatography-fluorescence detector for the determination\nof OTA in green coffee","volume":"310","author":"Moez","year":"2020","journal-title":"Food Chem."}],"container-title":["Analytical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.analchem.6c03182","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/38\/28158\/68492098\/acs.analchem.6c03182.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/ancham\/article-pdf\/98\/38\/28158\/68492098\/acs.analchem.6c03182.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T13:34:20Z","timestamp":1790688860000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/ancham\/article\/98\/38\/28158\/5428909\/Aptamer-Based-Sensing-with-Fluorescent-Light-Up"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,12]]},"references-count":43,"journal-issue":{"issue":"38","published-online":{"date-parts":[[2026,9,12]]},"published-print":{"date-parts":[[2026,9,29]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.analchem.6c03182","relation":{},"ISSN":["0003-2700","1520-6882"],"issn-type":[{"value":"0003-2700","type":"print"},{"value":"1520-6882","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,12]]}}}