BIOSENSORS & BIOELECTRONICS
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2025
·
IF 10.5
Bingjie Zheng,Yuqing Qin,Yanyan Chai,Aizhen Guo,Yonggang Hu
A DNA preamplification-free method was developed for sensitive detection of Salmonella typhimurium ( S. typhimurium ) by combining Pyrococcus furiosus Argonaute ( Pf Ago), a programmable small DNA-guided endonuclease and label-free DNAzyme, a functional nucleic acid (FNA). The effects of 5′-phosphorylated/hydroxylated single-strand DNA (5′P/OH ssDNA), including chimeric ssDNA-RNA and ssDNA-RNA-DNA (DNAzyme) on Pf Ago-mediated cleavage activity, were systematically investigated. Two detection strategies were developed: (1) A 63 nt 5′P DNAzyme was used as a guide DNA (gDNA) to mediate Pf Ago cleavage of probe MB1, resulting in a fluorescence signal inversely proportional to S. typhimurium concentration. RNase H2 from S. typhimurium (STH2) cleaved the DNAzyme, thereby reducing the signal. The limit of detection (LOD) was 4.5 × 10 CFU mL −1 , with a linear range from 8.0 × 10 to 6.4 × 10 4 CFU mL −1 (2) A 64 nt 5′OH DNAzyme was cleaved by STH2 to generate a 50 nt 5′P ssDNA, triggering Pf Ago-mediated MB2 cleavage. The fluorescence signal positively correlated with the concentration of S. typhimurium , with the LOD reduced to 2.0 × 10 CFU mL −1 and a detection range from 3.2 × 10 to 8.0 × 10 4 CFU mL −1 . Compared with the previous LOD of 3.2 × 10 3 CFU mL −1 achieved using the same DNAzyme labeled with a fluorescence probe and quencher, but without Pf Ago, the sensitivity of these methods was significantly improved, with increases of 71- and 160-folds, respectively. This method offers advantages of simplicity, specificity, rapidity, and sensitivity, making it suitable for point-of-care testing and holding promising potential for environmental monitoring, food safety, and clinical diagnosis.
研究领域:
分子诊断
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微生物检测
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核酸工程
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生物传感器
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食品安全
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2019
·
IF 9.518
Renyue Ji,Wuceng Niu,Shuai Chen,Wei Xu,Xingduo Ji,Lingyue Yuan,Hangyu Zhao,Minghao Geng,Jingfu Qiu,Chaorui Li
In this work, a novel Pb 2+ electrochemical DNAzyme sensor was developed for ultrasensitive detection of lead ions (Pb 2+ ) in water environment by coupling with the MoS 2 –AuPt nanomaterials and hemin/G-quadruplex DNAzyme, which acting as the electrocatalytic signal tag. Streptavidin (SA) modified tin dioxide-functionalized reduced graphene oxide (rGO-SnO 2 ) /gold nanoparticles (AuNPs) served as a sensor platform for enhancing conductivity and immobilizing more Pb 2+ -specific DNAzyme. In the presence of Pb 2+ , the Pb 2+ -dependent DNAzyme specifically reacted with Pb 2+ , cleaving the substrate strand (SS) into two free fragment and releasing the biotin-modified enzyme strand (Bio-ES) on the electrode. Connecting MoS 2 –AuPt nanocomposites labeled with G-rich DNA (G-DNA) strand and exposure of Bio-ES through the Helper DNA, as well as adding hemin to form a hemin/G-quadruplex, the biosensor achieved signal amplification . Chronoamperometry was used to record the current signal, which was primarily derived from the cocatalysis reduction of H 2 O 2 by MoS 2 –AuPt nanocomposites and the hemin/G-quadruplex. Under optimal conditions, the designed biosensor exhibited sensitive detection of Pb 2+ from 0.1 pg mL −1 to 1000 ng mL −1 , with a lower detection limit of 38 fg mL −1 (based on 3σ). This proposed biosensor is ultrasensitive and specific, representing a potential application for the detection of Pb 2+ in a water environment.
研究领域:
生物化学
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环境科学
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纳米技术
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分析化学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2014
·
IF 4.517
Chengke Wang,Xiaoya Dong,Qian Liu,Kun Wang
The combination of high selectivity of aptamer with the peroxidase-mimicking property of DNAzyme has presented considerable opportunities for designing colorimetric aptasensor for detection of ochratoxin A (OTA). The activities of both aptamer (as biorecognition element) and DNAzyme (as signal amplification element) are blocked via base pairing in the hairpin structure. Hybridization chain reaction (HCR) between two hairpin DNAs was employed to further improve the sensitivity of this method. The presence of OTA triggers the opening of the hairpin structure and the beginning of HCR, which results in the release of many DNAzyme, and generates enhanced colorimetric signals, which is correlated to the amounts of OTA with linear range between 0.01 to 0.32 nM, and the limit of detection is 0.01 nM under optimal conditions. OTA in yellow rice wine and wheat flour samples was also detected using this method. We demonstrate that a new colorimetric method for the detection of OTA has been established, which is simple, easy to conduct, label-free, sensitive, high throughput, and cost-saving.
研究领域:
食品科学
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分析化学
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生物技术
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DOI
PubMed
高影响因子
ACS Nano
·
2018
·
IF 13.709
Jie Feng,Zhen Xu,Feng Liu,Yun Zhao,Wenqian Yu,Min Pan,Fuan Wang,Xiaoqing Liu
Catalytic deoxyribozyme has great potential for gene regulation, but the poor efficiency of the cleavage of mRNA and the lack of versatile DNAzyme vehicles remain big challenges for potent gene therapy. By the rational designing of a diverse vehicle of polydopamine-Mn2+ nanoparticles (MnPDA), we demonstrate that MnPDA has integrated functions as an effective DNAzyme delivery vector, a self-generation source of DNAzyme cofactor for catalytic mRNA cleavage, and an inherent therapeutic photothermal agent as well as contrast agent for photoacoustic and magnetic resonance imaging. Specifically, the DNAzyme-MnPDA nanosystem protects catalytic deoxyribozyme from degradation and enhances cellular uptake efficiency. In the presence of intracellular glutathione, the nanoparticles are able to in situ generate free Mn2+ as a cofactor of DNAzyme to effectively trigger the catalytic cleavage of mRNA for gene silencing. In addition, the nanosystem shows high photothermal conversion efficiency and excellent stability against photothermal processing and degradation in complex environments. Unlike previous DNAzyme delivery vehicles, this vehicle exhibits diverse functionalities for potent gene regulation, allowing multimodal imaging-guided synergetic gene regulation and photothermal therapy both in vitro and in vivo.
研究领域:
生物化学
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分子生物学
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诊疗学
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纳米医学
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产品关联
DOI
PubMed
高影响因子
Communications Chemistry
·
2025
·
IF 6.2
Yang Xin,Fan Yiqi,Yao Bo,Shao Fangwei
G-quadruplex (GQ)/hemin DNAzymes represent a class of high-activity artificial peroxidases that have been widely utilized in various redox reactions. However, the limited thermal stability of GQ/hemin systems restricts their application in high-temperature biocatalysis. Herein, we developed a thermally activated peroxidase-mimetic DNAzyme constructed by G-quadruplex nanowire (GQwire) and hemin. A remarkable temperature-enhanced activity is shown on a model reaction of 2,2’-azino-bis(3-ethylben zothiozoline-6-sulfonic acid) (ABTS) oxidation by H 2 O 2 , achieving a 4.7-fold increase in catalytic activity and a 3.6-fold increase in cumulative product yield at 70 °C compared to 25°C. Structural analysis reveals that this unique thermal activation arises from the two-step thermal transition of GQwire architecture. GQwire disassembled from nanowire to oligomeric GQ rods that provides additional terminal binding sites for hemin and consequently enhancing catalytic activity at elevated temperatures. This thermal-activated DNAzyme provides an effective strategy for fabricating biocatalysts that expands their potentials in industrial manufacture.
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DOI
PubMed
高影响因子
Chemical Science
·
2025
·
IF 7.4
Baoting Dou,Hui Shen,Zhimin Li,Huanyu Cheng,Po Wang
Reactive oxygen species (ROS) play a critical role in regulating various physiological processes. To gain a comprehensive understanding of their distinct functions in different physiological events, it is imperative to detect binary ROS simultaneously. However, the development of the sensing method capable of binary ROS detection remains a significant challenge. In this study, we address this challenge by integrating chemically modified DNAzyme probes with the functionalized metal-organic framework (MOF) to create an efficient electrochemical sensing platform for the binary detection of ROS. ROS targets would activate the DNAzyme cleavage activity by removing the phenylboronate (BO) and phosphorothioate (PS) modifications, leading to the controlled release of doxorubicin (DOX) and methylene blue (MB) enclosed within MOF nanocomposites. This process generates two distinct voltammetric current peaks, with their potentials and intensities reflecting the identity and concentration of the ROS targets. The sensor demonstrates simultaneous detection of multiple ROS (H2O2 and HClO) produced by cancer cells with high sensitivity across a broad linear range of 1 to 200 nM and a low detection limit of sub-nanomolar. The design strategies behind the developed ROS sensing system could also be exploited to create other biosensors with highly sensitive and binary detection to promote clinical research and revolutionize disease diagnostics.
研究领域:
生物化学
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医学诊断
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Jie Xiang,Junyi Zhang,Shunmei Li,Ruo Yuan,Yun Xiang
The concentration variation of phenylalanine (Phe), an essential amino acid in humans, can cause metabolism disorders and even mental disability. Sensitive and convenient monitoring of Phe is therefore important for disease diagnosis. We describe here the establishment of a new aptamer-based, sensitive and label-free colorimetric Phe detection strategy by integrating catalytic hairpin assembly (CHA) and Mg 2+ -dependent DNAzyme amplification cascades. The target Phe coordinates with pentamethylcyclopentadienyl rhodium(III) chloride dimer [(Cp*RhCl 2 ) 2 ] to form a complex that has a high affinity to the corresponding aptamer sequence. Upon its binding to the aptamers in DNA duplex probes, ssDNA strands are released to trigger subsequent CHA reactions for the formation of many DNAzymes, which cleave the substrate signal probes to liberate lots of CHA initiation strands and free G-quadruplexes to realize the cascaded amplifications. Hemin further associates with the many G-quadruplexes to yield hemin/G-quadruplex mimicking peroxidases , which catalyze solution of substrate to exhibit highly enhanced UV–vis adsorption for detecting Phe at 0.19 μM level. At the meantime, the monitoring of Phe in diluted serums with high selectivity has also been demonstrated by the developed method, indicating its potential for simple diagnosis of Phe-related diseases.
研究领域:
生物化学
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医学诊断
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
ACS Sensors
·
2024
·
IF 8.2
Mengjie Chen,Zelin Yang,Zhuoliang Hu,Yudan Hao,Jing Lu,Duanping Sun
Monitoring biomarkers secreted by cardiomyocytes is critical to evaluate anticancer drug-induced myocardial injury (MI). Cardiac troponin I (cTnI) is considered the gold standard biomarker for MI. Herein, an electrochemical aptasensor is engineered for cTnI detection based on lanthanide europium metal–organic frameworks (Eu-MOFs) and a hybridization chain reaction-directed DNAzyme strategy. Three types of Eu-MOF morphologies were easily synthesized by changing the solvent, and the Eu-MOF modulated by mixing the solvent of dimethylformamide and H2O (D-Eu-MOF) exhibited the best performance compared to other morphologies of the Eu-MOFs. Multifunctional nanoprobes were constructed from D-Eu-MOF@Pt loaded with natural horseradish peroxidase and combined with an aptamer-initiated nuclear acid hybridization chain reaction to form G-quadruplex/hemin DNAzymes for signal amplification. A novel capture probe is constructed on the basis of DNA nanotetrahedrons modified on screen-printed gold electrodes to enhance the capture of the target and multifunctional nanoprobes for signal amplification. It exhibits a detection limit of 0.17 pg mL–1 and a linear range from 0.5 pg mL–1 to 15 ng mL–1. The practicality of the platform is evaluated by measuring cTnI in real samples and secreted by cardiomyocytes after drug treatment, which provides great potential in drug-induced MI evaluation for clinical application.
研究领域:
生物医学工程
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分子诊断学
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DOI
PubMed
高影响因子
Science China-Chemistry
·
2019
·Vol. 63
·期 3
·404-410
·
IF 6.085
Lu Shasha,Wang Shuang,Zhao Jiahui,Sun Jian,Yang Xiurong
RNA-cleaving DNAzymes possess important roles in DNAzymes and have been widely used in the biosensors, DNA nanomachines owing to their ion-specific dependence. However, there are still challenges in constructing universal but versatile stimuli-responsive strategies of RNA-cleaving DNAzymes. Herein, a stimuli-responsive strategy for RNA-cleaving DNAzyme is proposed by the artful design of hairpin nanostructure, in which the activities of DNAzyme (Pb 2+ -dependent DNAzyme as a model) in the hairpin’s loop are pH-regulated by using the triplex stem as the “lock”. Upon introducing the “key”, pH values, the DNAzyme will be activated and fragment the substrate of it in the presence of Pb 2+ , accompanied by the turn-on of the fluorescence quenched by fluorescence resonance energy transfer (FRET). The regulation ability of pH can be controlled by the length and sequence of the triplex stem, and the wide pH regulation range may be helpful for the application of DNAzymes in biological medicine delivery systems.
研究领域:
生物化学
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纳米技术
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生物传感器
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分子生物学
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产品关联
DOI
高影响因子
Bioactive Materials
·
2022
·
IF 16.874
Yi Xiao,Haomin Yi,Jingzhi Zhu,Suhua Chen,Guofang Wang,Yilong Liao,Yuanyuan Lei,Liyin Chen,Xingcai Zhang,Fangfu Ye
Toxicity assessment is a major problem in pharmaceutical candidates and industry chemicals development. However, due to the lack of practical analytical methods for DNA adduct analysis, the safety evaluation of drug and industry chemicals was severely limited. Here, we develop a DNAzyme-based method to detect DNA adduct damage for toxicity assessment of drugs and chemicals. Among 18 structural variants of G4 DNAzyme, EA2 DNAzyme exhibits an obvious DNA damaging effect of styrene oxide (SO) due to its unstable structure. The covalent binding of SO to DNAzyme disrupts the Hoogsteen hydrogen bonding sites of G-plane guanines and affects the formation of the G4 quadruplex. DNA damage chemicals reduce the peroxidase activity of the G4 DNAzyme to monitor the DNA adduct damage by disrupting the structural integrity of the G4 DNAzyme. Our method for genotoxic assessment of pharmaceutical candidates and industrial chemicals can elucidate the complex chemical pathways leading to toxicity, predict toxic effects of chemicals, and evaluate possible risks to human health.
研究领域:
分子生物学
·
毒理学
·
药学
·
化学工程
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产品关联
DOI
PubMed
高影响因子
Chemosensors
·
2022
·Vol. 10
·期 7
·276
·
IF 4.229
Kunhan Nie,Yongjian Jiang,Na Wang,Yajun Wang,Di Li,Lei Zhan,Chengzhi Huang,Chunmei Li
The abnormal expression of miRNA is closely related to the occurrence of pancreatic cancer. Herein, a programmable DNAzyme amplifier for the universal detection of pancreatic cancer-related miRNAs was proposed based on its programmability through the rational design of sequences. The fluorescence signal recovery of the DNAzyme amplifier showed a good linear relationship with the concentration of miR-10b in the range of 10–60 nM, with a detection limit of 893 pM. At the same time, this method displayed a high selectivity for miR-10b, with a remarkable discrimination of a single nucleotide difference. Furthermore, this method was also successfully used to detect miR-21 in the range of 10–60 nM based on the programmability of the DNA amplifier, exhibiting the universal application feasibility of this design. Overall, the proposed programmable DNAzyme cycle amplifier strategy shows promising potential for the simple, rapid, and universal detection of pancreatic cancer-related miRNAs, which is significant for improving the accuracy of pancreatic cancer diagnosis.Keywords:DNAzyme amplifier;signal amplification;miRNA detection;pancreatic cancer
研究领域:
分子生物学
·
医学诊断
·
生物化学
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产品关联
DOI
高影响因子
TALANTA
·
2023
·
IF 6.1
Jingyi Dong,Xia Qiu,Mimi Huang,Xiaohu Chen,Yongxin Li
Solid-nanopores/nanopipettes have the exquisite ability to reveal the changes in molecular volume due to the advantages of adjustable size, good rigidity and low noise. Herein, a new platform for sensing application was established based on G-quadruplex-hemin DNAzyme (GQH) functionalized gold-coated nanopipettes. In this method, GQH was immobilized on gold-coated nanopipette, which could be used as a catalyst for the reaction of H 2 O 2 with 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) to promote the conversion of ABTS to ABTS + ions inside gold-coated nanopipette, and the change of transmembrane ion current could be monitored in real time. At the optimal conditions, there was a correlation between the ion current and the concentration of H 2 O 2 in a certain range, which could be used for the hydrogen peroxide sensing. The GQH immobilized nanopipette provides a useful platform to investigate enzymatic catalysis in confined environment, which can be used in electrocatalysis , sensing and fundamental electrochemistry .
研究领域:
纳米技术
·
生物化学
·
分析化学
·
电化学
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产品关联
DOI
PubMed
高影响因子
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