Target triggered ultrasensitive electrochemical polychlorinated biphenyl aptasensor based on DNA microcapsules and nonlinear hybridization chain reaction.

In this work, we demonstrated an ultrasensitive detection platform for polychlorinated biphenyls (PCBs) based on DNA microcapsules and a nonlinear hybridization chain reaction (NHCR). In the process, first, electrochemical signal molecules (Methylene Blue, MB) were sealed in the prepared DNA microcapsules. In the presence of PCB-72, DNA microcapsules could be dissociated with the conjugation of the aptamer and target, and meanwhile, the released DNA strand could initiate the NHCR and trigger the chain branching growth of DNA dendrimers. Because the released MBs were intercalated into the DNA dendrimer, enhanced electrochemical responses could be detected. This method exhibited ultrahigh sensitivity to PCB-72 with a detection limit of 0.001 ng mL-1. Furthermore, the present aptasensor was also capable of discriminating different PCB congeners. Therefore, the devised label-free and enzyme-free amplification electrochemical aptasensor strategy has great potential for the detection of PCB-72 in real samples, and this strategy may also become an attractive alternative for sensitive and selective small molecule, protein, nucleic acid and nuclease activity detection.

Autonomous operation of 3D DNA walkers in living cells for microRNA imaging.

Three dimensional (3D) DNA walkers hold great potential in serving as an ideal candidate for signal transduction and amplification in bio-assays. However, the autonomous operation of 3D DNA walkers inside living cells is still few and far between, which could be attributed to the lack of suitable driving forces and moderate efficiency in terms of the cellular uptake of such complex 3D DNA components. Herein, a newly updated autonomously operated and highly integrated 3D DNA walker on Au nanoparticles (Au NPs)/zeolitic imidazolate framework-8 (ZIF-8) was activated in a tumor microenviroment and its signal amplified assay capability in living cells was demonstrated using miRNA as a sensing model biomolecule. Specifically, we assembled a 3D DNA motor, including Zn2+-dependent DNAzyme and substrates on the AuNPs grafted on ZIF-8. After being delivered into a living cell, ZIF-8 was efficiently degraded in the tumor microenvironment (low pH value), locally releasing the Zn2+ and DNA motor. Then, a self-sufficient DNA motor autonomously performed the bio-analytical task of imaging miRNA-10b, with a low detection limit of 34 pM. Also, such self-sufficient 3D walkers allowed real-time imaging of MDA-MB-231 cells by intracellular operation. This method demonstrates the self-sufficient 3D DNA motor's bioanalytical application in living cells which may inspire various other biological applications including gene delivery, therapy, etc.

In Situ Cation Exchange Generated ZnS-Ag2S Nanoparticles for Photothermal Detection of Transcription Factor.

The photothermal reagent (PTA)-mediated point-of-care detection of disease biomarkers using thermometers as readout has attracted increasing attention, but complex modification or generation process of PTAs still limited their further applications. Herein, we report a photothermal detection platform in which the target recognition triggered the in situ generation of the PTA and ZnS-Ag2S nanoparticles (NPs) by one-step autonomous cation exchange reaction (ACER). As a proof of concept, NF-kB1, a kind of disease biomarker, was used to demonstrate the photothermal detection platform. First, a triplex-like DNA structure containing the recognition part of NF-kB1 and cytosine-Ag+-cytosine (C-Ag+-C) unit was designed. With the recognition of NF-kB1, abundant C-Ag+-C units were released to take ACER with the prepared ZnS NPs. In addition, because of the intrinsic detecting limitation of the thermometer, hybridization chain reaction (HCR) was introduced to load more Ag+ for the enhancement of the photothermal signal. After the irradiation of 808 nm laser toward the generated ZnS-Ag2S NPs dispersions, its temperature increased with the increased concentration of NF-kB1 in the range from 10 to 1000 nM with a detection limit of 2.31 nM. The signal-amplifying role of HCR was confirmed by the control experiment. This platform of in situ generation of PTA not only integrated the recognition process with detection but also avoided the complex process of modification or generation, which has remarkable potential to be extended for the detections of other different disease biomarkers.

Plasmon-Enhanced Electrochemiluminescence of Silver Nanoclusters for microRNA Detection.

Surface plasmon-enhanced electrochemiluminescence (SPEECL) with excellent sensitivity and simplicity has attracted increasing attention. In this work, we reported a novel SPEECL with DNA templated silver nanoclusters (DNA-AgNCs) as ECL emitters and gold nanoparticles (AuNPs) as localized surface plasmon resonance (LSPR) source. The SPEECL with DNA-AgNCs as ECL luminophores possessed low toxicity and avoided the labeling process, which is favorable for its further sensing application. In addition, by investigation of the SPEECL under different distances between DNA-AgNCs and AuNPs, it was demonstrated that the SPEECL was distance dependent. Meanwhile, the SPEECL intensity changed with the sizes and interdistance of AuNPs under different electrodeposition time. Furthermore, by the combination of a cyclic amplification process with enzyme-free catalytic hairpin DNA, a sensitive SPEECL biosensor was proposed for the detection of microRNA (miRNA-21) successfully with a wide linear range from 1 aM to 104 fM and a relatively low detection limit of 0.96 aM, which was applied in the detection of miRNA-21 in real samples with satisfying results. This novel, simple, sensitive, and selective SPEECL with label-free and low-toxic ECL emitters displayed a great potential for bioassay application.