Imaging Endogenous Metal Ions in Living Cells Using a DNAzyme–Catalytic Hairpin Assembly Probe

Wu, Zhenkun; Fan, Huanhuan; Satyavolu, Nitya Sai Reddy; Wang, Wenjing; Lake, Ryan J.; Jiang, Jian‐Hui; Lu, Yi

doi:10.1002/ange.201703540

Cited by 52 publications

(28 citation statements)

References 65 publications

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“…As a versatile SDA isothermal amplification technique, catalytic hairpin assembly (CHA) is an enzyme-free DNA circuit that relies on a thermodynamically driven entropy gain process to achieve signal amplification, possibly permitting exponential amplification of a target sequence in 15 min 34 - 36 . By combining CHA with other detection techniques such as colorimetry 37 , fluorescence 38 , electrochemistry 39 , 40 , chemiluminescence 41 , Raman spectroscopy, and surface plasmon resonance, miscellaneous CHAs platforms have been developed for miRNAs, metal ions 42 and protein molecular analysis 43 . However, these strategies are associated with complex nanoparticles and fluorescence labelling processes, time-consuming methods or expensive equipment.…”

Section: Introductionmentioning

confidence: 99%

Catalytic hairpin assembly gel assay for multiple and sensitive microRNA detection

et al. 2018

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As important modulators of gene expression, microRNAs (miRNAs) have been identified as promising biomarkers with powerful predictive value in diagnosis and prognosis for several diseases, especially for cancers. Here we report a facile, multiple and sensitive miRNA detection method that uses conventional gel electrophoresis and catalytic hairpin assembly (CHA) system without any complex nanomaterials or enzymatic amplification.Methods: In this study, three pairs of hairpin probes are rationally designed with thermodynamically and kinetically preferable feasibility for the CHA process. In the present of target miRNA, the stem of the corresponding hairpin detection probe (HDP) will be unfolded and expose the concealed domain. The corresponding hairpin assistant probe (HAP) then replaces the hybridized target miRNA to form specific HDP/HAP complexes and releases miRNA based on thermodynamically driven entropy gain process, and the released miRNA triggers the next recycle to produce tremendous corresponding HDP/HAP complexes.Results: The results showed that the CHA gel assay can detect miRNA at fM levels and shows good capability of discriminating miRNA family members and base-mismatched miRNAs. It is able to analyze miRNAs extracted from cell lysates, which are consistent with the results of conventional polymerase chain reaction (PCR) method. Depending on the length of the designed hairpin probes, the CHA gel assay consisting of different hairpin probes effectively discriminated and simultaneously detected multiple miRNAs in homogenous solution and miRNAs extracted from cell lysates.Conclusion: The work highlights the practical use of a conventional gel electrophoresis for sensitive interesting nucleic acid sequences detection.

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Section: Introductionmentioning

confidence: 99%

Catalytic hairpin assembly gel assay for multiple and sensitive microRNA detection

et al. 2018

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“…The RCA product was detected by a duplex-binding dye (i.e., 3, 3 0 -diethylthiadicarbocyanine), exhibiting a detection limit as low as 10 cells/mL. In another example, Wu et al utilized the CHA reaction to enhance the sensitivity of the Na + -dependent DNAzyme (NaA43) (Wu et al, 2017). In the presence of Na + , the cleavage substrate fragment (red) initiates the CHA reaction by opening the hairpin DNA, H1 ( Figure 10B).…”

Section: Applications In Signal Amplification Smart Materials Assembmentioning

confidence: 99%

Catalytic Nucleic Acids: Biochemistry, Chemical Biology, Biosensors, and Nanotechnology

Liu

2020

iScience

131

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Since the initial discovery of ribozymes in the early 1980s, catalytic nucleic acids have been used in different areas. Compared with protein enzymes, catalytic nucleic acids are programmable in structure, easy to modify, and more stable especially for DNA. We take a historic view to summarize a few main interdisciplinary areas of research on nucleic acid enzymes that may have broader impacts. Early efforts on ribozymes in the 1980s have broken the notion that all enzymes are proteins, supplying new evidence for the RNA world hypothesis. In 1994, the first catalytic DNA (DNAzyme) was reported. Since 2000, the biosensor applications of DNAzymes have emerged and DNAzymes are particularly useful for detecting metal ions, a challenging task for enzymes and antibodies. Combined with nanotechnology, DNAzymes are key building elements for switches allowing dynamic control of materials assembly. The search for new DNAzymes and ribozymes is facilitated by developments in DNA sequencing and computational algorithms, further broadening our fundamental understanding of their biochemistry.

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“…Among them, the electrochemical aptasensor is the most extensively used because of its operational feasibility and sensitivity [15][16]. In addition, different signal amplification techniques such as rolling circle amplification (RCA) [17][18], hybridization chain reaction (HCR) [19][20] and catalytic hairpin assembly (CHA) [21][22] are also incorporated with these detection methods to further improve detection sensitivity. Interestingly, CHA has higher catalytic efficiency, lower background signal, simpler and more stable reaction system [23][24].…”

Section: Introductionmentioning

confidence: 99%

Enzyme‐free Recycling Amplification‐based Sensitive Electrochemical Thrombin Aptasensor

Zhang¹,

Liao²,

Wang³

et al. 2021

Electroanalysis

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In this work, an enzyme‐free recycling amplification‐based sensitive electrochemical thrombin aptasensor using catalytic hairpin assembly (CHA) and bisferrocene was proposed for electrochemical beacon. Thrombin triggered the CHA reaction that in turn formed a number of double‐stranded complexes,which are fixed on the surface of the gold electrode through potential‐assisted Au−S deposition, and a large number of labeled bisferrocene are placed close to the gold electrode to generate electrochemical signal. The linear range of the electrochemical sensor was 0.25 pM‐2.5 nM for thrombin with detection limit of 0.18 pM. This sensor can be employed to monitor the thrombin in human serum samples.

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Imaging Endogenous Metal Ions in Living Cells Using a DNAzyme–Catalytic Hairpin Assembly Probe

Abstract: Supportinginformation and the ORCID identification number(s) for the author(s) of this article can be found under: https://doi.

Cited by 52 publications

References 65 publications

Catalytic hairpin assembly gel assay for multiple and sensitive microRNA detection

Catalytic hairpin assembly gel assay for multiple and sensitive microRNA detection

Catalytic Nucleic Acids: Biochemistry, Chemical Biology, Biosensors, and Nanotechnology

Enzyme‐free Recycling Amplification‐based Sensitive Electrochemical Thrombin Aptasensor

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