title: Synthesis and sensing integration: A novel enzymatic reaction modulated Nanoclusters Beacon (NCB) "Illumination" strategy for label-free biosensing and logic gate operation.
abstract: A novel fluorescent label-free "turn-on" NAD(+) and adenosine triphosphate (ATP) biosensing strategy is proposed by fully exploiting ligation triggered Nanocluster Beacon (NCB). In the presence of the target, the split NCB was brought to intact, which brought the C-rich sequence and enhancer sequence in close proximity resulting in the lightening of dark DNA/AgNCs ("On" mode). Further application was presented for logic gate operation and aptasensor construction. The feasibility was investigated by Ultraviolet-visible spectroscopy (UV-vis), Fluorescence, lifetime and High Resolution Transmission Electron Microscopy (HRTEM) etc. The strategy displayed good performance in the detection of NAD(+) and ATP, with the detection limit of 0.002nM and 0.001mM, the linear range of 10-1000nM and 0.003-0.01mM, respectively. Due to the DNA/AgNCs as fluorescence reporter, the completely label-free fluorescent strategy boasts the features of simplicity and low cost, and showing little reliance on the sensing environment. Meanwhile, the regulation by overhang G-rich sequence not relying on Förster energy transfer quenching manifests the high signal-to-background ratios (S/B ratios). This method not only provided a simple, economical and reliable fluorescent NAD(+) assay but also explored a flexible G-rich sequence regulated NCB probe for the fluorescent biosensors. Furthermore, this sensing mode was expanded to the application of a logic gate design, which exhibited a high performance for not only versatile biosensors construction but also for molecular computing application.
authors.affiliation: Key Laboratory of Functional Molecular Solids, Wuhu, Anhui Province, 241000 PR China; College of Chemistry and Materials Science, Center for Nano Science and Technology, Anhui Normal University, Wuhu, 241000 PR China; Key Laboratory of Chem-Biosensing, Wuhu, Anhui Province, 241000 PR China.
authors.forename: Lu
authors.initials: L
authors.lastname: Hong
authors.affiliation: Key Laboratory of Functional Molecular Solids, Wuhu, Anhui Province, 241000 PR China; College of Chemistry and Materials Science, Center for Nano Science and Technology, Anhui Normal University, Wuhu, 241000 PR China; Key Laboratory of Chem-Biosensing, Wuhu, Anhui Province, 241000 PR China.
authors.forename: Fu
authors.initials: F
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Functional Molecular Solids, Wuhu, Anhui Province, 241000 PR China; College of Chemistry and Materials Science, Center for Nano Science and Technology, Anhui Normal University, Wuhu, 241000 PR China; Key Laboratory of Chem-Biosensing, Wuhu, Anhui Province, 241000 PR China; State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha, 410082 PR China. Electronic address: wangyuz@mail.ahnu.edu.cn.
authors.forename: Guangfeng
authors.initials: G
authors.lastname: Wang
authors.affiliation: Key Laboratory of Functional Molecular Solids, Wuhu, Anhui Province, 241000 PR China; College of Chemistry and Materials Science, Center for Nano Science and Technology, Anhui Normal University, Wuhu, 241000 PR China; Key Laboratory of Chem-Biosensing, Wuhu, Anhui Province, 241000 PR China.
authors.forename: Xiaojun
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis*; D039301:Computers, Molecular; D004247:DNA / Q000737:chemistry* / Q000528:radiation effects; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D008029:Lighting / Q000295:instrumentation; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D009243:NAD / Q000032:analysis*; D058726:Nanoconjugates / Q000737:chemistry* / Q000528:radiation effects; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012815:Signal Processing, Computer-Assisted / Q000295:instrumentation*; D012834:Silver / Q000737:chemistry; D013194:Staining and Labeling
chemical_list: D058726:Nanoconjugates; D009243:NAD; D012834:Silver; D000255:Adenosine Triphosphate; D004247:DNA
keywords: Ag nanocluster; Label-free; Logic gate; NAD(+); Nanoclusters Beacon (NCB)