title: Dual-color encoded DNAzyme nanostructures for multiplexed detection of intracellular metal ions in living cells.
abstract: The detection of intracellular metal ions is of great importance in understanding metal homeostasis in cells and related diseases, and yet it remains a significant challenge to achieve this goal. Based on a new self-assembled and dual-color encoded DNAzyme nanostructure, we describe here an approach for multiplexed sensing of UO2(2+) and Pb(2+) in living cells. The fluorescently quenched nanoprobes can be prepared by simple thermal annealing of four ssDNAs containing the metal ion-dependent enzymatic and substrate sequences. The self-assembly formation of the nanostructures are verified by native polyacrylamide gel electrophoresis. The target metal ions can cleave the substrate sequences in the DNAzyme nanostructures to recover fluorescent emissions at different wavelengths for sensitive and selective in vitro multiplexed detection of UO2(2+) and Pb(2+) with the detection limits of 0.6nM and 3.9nM, respectively. Importantly, we demonstrate that these nanoprobes are stable in cell lysates and can enter cells without the aid of any transfection agents for simultaneous imaging intracellular UO2(2+) and Pb(2+). Moreover, the nanoprobes offer excellent biocompatibility and non-cytotoxicity. With these unique features, the dual-color encoded nanostructures presented here can thus offer new opportunities for multiplexed detection of specific intracellular species.
authors.affiliation: Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
authors.forename: Wenjiao
authors.initials: W
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
authors.forename: Wenbing
authors.initials: W
authors.lastname: Liang
authors.affiliation: Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
authors.forename: Daxiu
authors.initials: D
authors.lastname: Li
authors.affiliation: Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
authors.forename: Ruo
authors.initials: R
authors.lastname: Yuan
authors.affiliation: Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China. Electronic address: yunatswu@swu.edu.cn.
authors.forename: Yun
authors.initials: Y
authors.lastname: Xiang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D002413:Cations, Divalent / Q000032:analysis; D002470:Cell Survival; D021881:DNA, Catalytic / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D007854:Lead / Q000032:analysis*; D018613:Microscopy, Confocal / Q000379:methods; D049329:Nanostructures / Q000737:chemistry*; D061848:Optical Imaging / Q000379:methods*; D013050:Spectrometry, Fluorescence / Q000379:methods; D017974:Uranium Compounds / Q000032:analysis*
chemical_list: D002413:Cations, Divalent; D021881:DNA, Catalytic; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D017974:Uranium Compounds; D007854:Lead; C012597:uranium dioxide
keywords: DNAzyme nanostructure; Intracellular imaging; Metal ions; Multiplexed detection