TALANTA
·
2016
·
IF 4.035
Lu Hong,Jing Wan,Xiaojun Zhang,Guangfeng Wang
In this work, a highly sensitive electrochemical DNA methyltransferase (MTase) activity assay was fabricated with DNA–gold nanoparticles (Au NPs) network as signal amplification unit and an easy assembly method by the linkage of benzenedithiol bridge. By two complementary AuNPs modified single-stranded DNA, DNA–gold nanoparticles network was self-assembled. With the linkage of benzenedithiol bridge, the DNA network structure was immobilized on the surface of gold electrode through the covalent Au–S bond. In the presence of Dam MTase, the special sites of DNA–AuNPs network were methylated and could not be digested by restriction endonuclease Mbo I. Thus the loaded electrochemical indicator Methylene blue (MB) was MB molecules still remained on the DNA–Au NPs network. The electrochemical response depended on the methylated degree, which could be used to detect MTase activity. By the differential pulse voltammetry (DPV), it was demonstrated that a linear relationship between the DPV response and logarithm of Dam concentration ranged from 0.075 to 30 U/mL, achieving a low detection limit of 0.02 U/mL. The use of benzenedithiol avoided the direct incubation of the solid electrode with the capture DNA probe under complex and harsh conditions. Therefore the immobilization of DNA–AuNPs network was easy to be carried out, which is favorable for the specially high stability and reproducibility of the electrochemical biosensor.
研究领域:
生物化学
·
分子生物学
·
纳米技术
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR LIQUIDS
·
2019
·
IF 4.561
Rui Wang,Xing Hu,Junhui Pan,Guowen Zhang,Deming Gong
Isoeugenol , a natural phenolic compound in plant essential oils, is known to have many biological and pharmacological properties . In the present work, the interaction of isoeugenol with calf thymus DNA (ctDNA) and its protective effect on DNA oxidative damage were investigated by multispectroscopic techniques, viscosity measurements, gel electrophoresis studies and molecular simulation. Analyses of ultraviolet-visible (UV–vis) absorption and fluorescence spectra indicated that isoeugenol could bind to ctDNA with a binding constant on the order of 10 3 L mol −1 , which was mainly driven by hydrogen bonding and van der Waals forces . The results from the viscosity measurements, competitive displacement studies and circular dichroism (CD) analysis indicated that the binding of isoeugenol to ctDNA was in the groove mode and that the A–T base pairs in the minor grooves of ctDNA were the main binding sites for isoeugenol, which was supported by molecular docking analyses. In addition, isoeugenol was found to show a protective effect against hydroxyl-induced DNA damage, which may be explained by its good antioxidant activity . These findings may improve the understanding of the mechanism of interaction between isoeugenol and ctDNA and provide a theoretical basis for the development of isoeugenol in the food and drug industry.
研究领域:
药理学
·
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 3
·1-7
·
IF 5.7
Li Xiang-Ling,Jiang Han,Zhao Lei,Song Tian shun,Xie Jing jing
A versatile DNA nanomachine detection system has been developed via the combination of DNAzyme with catalytic hairpin assembly (CHA) technology for achieving accurate and sensitive detection of lead ions (Pb 2+ ). In the presence of target Pb 2+ , capture DNA nanomachine formed by AuNP and DNAzyme recognized and reacted with Pb 2+ , which yielded an “active” DNAzyme, that induced the cleavage of substrate strand, and then released the initiator DNA (TT) for CHA. With the help of the initiator DNA TT, self-powered CHA was activated to achieve the signal amplification reaction in the detection of DNA nanomachine. Meanwhile, the initiator DNA TT was released and hybridized with the other H1 strand to initiate another CHA, replacement, and turnovers, producing enhanced fluorescence signal of fluorophore FAM (excitation 490 nm/emission 520 nm) for sensitive determination of Pb 2+ . Under the optimized conditions, the DNA nanomachine detection system revealed high selectivity toward Pb 2+ in the concentration range 50–600 pM, with the limit of detection (LOD) of 31 pM. Recovery tests demonstrated that the DNA nanomachine detection system has excellent detection capability in real samples. Therefore, the proposed strategy can be extended and act as a basic platform for highly accurate and sensitive detection of various heavy metal ions. Graphical Abstract
研究领域:
纳米技术
·
分析化学
·
生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2018
·
IF 4.244
Xiaojuan Liu,Ningning Xu,Panpan Gai,Feng Li
Since melamine is a strong hazard to human health, the development of new methods for highly sensitive detection of melamine is highly desirable. Herein, a novel fluorescent biosensing strategy was designed for sensitive and selective melamine assay based on the recognition ability of abasic (AP) site in triplex towards melamine and signal amplification by Mg 2+ -dependent DNAzyme. In this strategy, the melamine-induced formation of triplex DNA was employed to trigger the strand displacement reaction (SDR). The SDR process converted the specific target recognition into the release and activation of Mg 2+ -dependent DNAzyme, which could catalyze the cleavage of fluorophore/quencher labeled DNA substrate (FQ), resulting in a significantly increased fluorescent signal. Under the optimal conditions, the fluorescent signal has a linear relationship with the logarithm of the melamine concentration in a wide range of 0.005–50 μM. The detection limit was estimated to be 0.9 nM (0.1ppb), which is sufficiently sensitive for practical application. Furthermore, this strategy exhibits high selectivity against other potential interfering substances, and the practical application of this strategy for milk samples reveals that the proposed strategy works well for melamine assay in real samples. Therefore, this strategy presents a new method for the sensitive melamine assay and holds great promise for sensing applications in the environment and the food safety field.
研究领域:
生物化学
·
食品安全
·
分析化学
✓ 已验证
DOI
高影响因子
LANGMUIR
·
2018
·
IF 3.789
Alessio Caciagli,Mykolas Zupkauskas,Aviad Levin,Tuomas P. J. Knowles,Clément Mugemana,Nico Bruns,Thomas O’Neill,William J. Frith,Erika Eiser
Many industrial soft materials include oil-in-water (O/W) emulsions at the core of their formulations. By using tuneable interface stabilizing agents, such emulsions can self-assemble into complex structures. DNA has been used for decades as a thermoresponsive, highly specific binding agent between hard and, recently, soft colloids. Up until now, emulsion droplets functionalized with DNA had relatively low coating densities and were expensive to scale up. Here, a general O/W DNA-coating method using functional nonionic amphiphilic block copolymers, both diblock and triblock, is presented. The hydrophilic poly(ethylene glycol) ends of the surfactants are functionalized with azides, allowing for efficient, dense, and controlled coupling of dibenzocyclooctane-functionalized DNA to the polymers through a strain-promoted alkyne–azide click reaction. The protocol is readily scalable due to the triblock’s commercial availability. Different production methods (ultrasonication, microfluidics, and membrane emulsification) are used with different oils (hexadecane and silicone oil) to produce functional droplets in various size ranges (submicron, ∼20 and >50 μm), showcasing the generality of the protocol. Thermoreversible submicron emulsion gels, hierarchical “raspberry” droplets, and controlled droplet release from a flat DNA-coated surface are demonstrated. The emulsion stability and polydispersity is evaluated using dynamic light scattering and optical microscopy. The generality and simplicity of the method opens up new applications in soft matter, biotechnological research, and industrial advances.
研究领域:
软物质科学
·
材料科学
·
生物技术
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Hui-Fang Cui,Tai-Bin Xu,Yu-Long Sun,An-Wei Zhou,Yu-Han Cui,Wei Liu,John H. T. Luong
Hairpin DNA (hpDNA) as a novel biobarcode was conjugated with gold nanoparticles (AuNPs) and a reporter DNA (rpDNA) to form hpDNA/AuNP/rpDNA nanoparticles for the detection of an oligonucleotide sequence associated with Helicobacter pylori as a model target. The rpDNA is complementary to about a half-portion of the target DNA sequence (tDNA). A capture DNA probe (cpDNA), complementary to the other half of the tDNA, was immobilized on the surface of a gold electrode. In the presence of tDNA, a sandwich structure of (hpDNA/AuNP/rpDNA)/tDNA/cpDNA was formed on the electrode surface. The differential pulse voltammetry (DPV) detection was based on [Ru(NH3)5(3-(2-phenanthren-9-yl-vinyl)-pyridine)]2+, an electroactive complex that binds to the sandwich structure by its intercalation with the hpDNA and the double-stranded DNA (dsDNA) of the sandwich structure. The several factors—high density of biobarcode hpDNA on the surface of AuNPs, multiple electroactive complex molecules intercalated with each hpDNA and dsDNA molecule, and the intercalation binding mode of the electroactive complex with the DNA sandwich structure—contribute to the DNA sensor with highly selective and sensitive sensing properties. The DNA sensor exhibited a detection limit of 1 × 10–15 M (i.e., 1 fM), the DNA levels in physiological samples, with linearity down to 2 × 10–15 M. It can differentiate even one single mismatched DNA from the complementary tDNA. This novel biobarcode-based DNA sensing approach should provide a general platform for development of direct, simple, repetitive, sensitive, and selective DNA sensors for various important applications in analytical, environmental, and clinical chemistry.
研究领域:
生物传感器
·
分析化学
·
临床医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Dawei Li,Shen Ling,Hongsong Wu,Zhaoqi Yang,Bei Lv
In addition to its application in genome editing, the CRISPR/Cas system provides a new molecular tool for the construction of biosensor platforms due to its high target sequence specificity and programmability . To exploit the potential of the CRISPR/Cas system for detecting non-nucleic-acid targets, two ultrasensitive biosensors (sensor-ss and sensor-ds) consisting of allosteric aptamer probes integrated with the CRISPR/Cas12a system were designed and constructed for tobramycin detection. Both sensors utilized hairpin DNA containing a tobramycin aptamer sequence as the target recognition probe, which can respond to target binding and further produce signal transduction sequences. However, there are two differences between the sensors. First, in sensor-ss, single-stranded DNA was set to be the activator to trigger cutting activity ( trans -cleavage) of CRISPR/Cas12a, leading to cleavage of the FQ-reporter (a short single-stranded DNA labeled with a fluorophore and quencher at each end) and increasing the fluorescent signal. However, double-stranded DNA containing a protospacer adjacent motif (PAM) site was produced in sensor-ds as the signal transduction sequence, which was recognized by CRISPR/Cas12a, resulting in the cleavage of the FQ-reporter. Second, sensor-ss involved a strand displacement amplification (SDA) reaction as the signal enhancement step, while no DNA amplification procedure was designed in sensor-ds. Through DNA sequence and reaction condition optimization, sensor-ds showed a linear relationship between the fluorescence response and tobramycin concentrations ranging from 10 to 300 pM with a limit of detection (LOD) as low as 3.719 pM. Although sensor-ss exhibited higher sensitivity (LOD = 1.542 pM) than sensor-ds, rapidly reaching the fluorescent signal saturation platform led to a narrow linear response range (5–30 pM). Moreover, our CRISPR/Cas12a-based assay can realize on-site detection by the naked eye readout under UV light. The constructed sensor-ds was also used to detect tobramycin in milk and lake water samples, and the results indicated that the detection system was reliable. The design strategy reported in this study could shed new light on CRISPR/Cas12a-based biosensor construction for small molecule detection.
研究领域:
分子生物学
·
分析化学
·
医学检测
✓ 已验证
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Boyuan Jia,Xuesong Li,Xinyu Wang,Yanyan Lu,Hui Meng,Xinshuo Zhang,Xiangyang Wu,Zhen Zhang,Yibin Deng
DNA-templated silver nanoclusters (DNA-AgNCs) exhibit infinite application foreground for biosensing due to their fluorescence enhancement properties induced by G-quadruplex (G4). Albeit advanced, their synergistic effects of G4 topologies, and spatial proximity between G4 and DNA-AgNCs remain unclear, which will influence the design of DNA-AgNCs based biosensors and their performance. Herein, the fluorescence enhancement of DNA-AgNCs initiated by three distinct G4 topologies (parallel antiparallel, hybrid) and their distance-dependent behaviors were systematically investigated, using a DNA tetrahedron as a mechanically rigid scaffold to precisely regulate the distance between DNA-AgNCs and G4. We found that parallel G4 structures significantly boosted the fluorescence intensity of DNA-AgNCs, and their closer proximity resulted in elevated fluorescence, which were ascribed to a denser surrounding electron environment that facilitates electron transfer to nearby AgNCs, achieving fluorescence enhancement. Consequently, after encoding a specific aptamer into the reconfigurable edge of the DNA tetrahedron, a novel biosensor was fabricated as a proof-of-concept for rapid detection of aflatoxin B1 with a detection limit of 0.068 ng/mL. Our strategy offered some new insights for rational bioassay design and its performance improvement.
研究领域:
生物化学
·
纳米生物学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
Nature Communications
·
2022
·Vol. 13
·期 1
·1-14
·
IF 17.694
Zhou Liang,Lei Qi,Guo Jimin,Gao Yuanyuan,Shi Jianjun,Yu Hong,Yin Wenxiang,Cao Jiangfan,Xiao Botao,Andreo Jacopo,Ettlinger Romy,Jeffrey Brinker C.,Wuttke Stefan,Zhu Wei
Deoxyribonucleic acid (DNA) is the blueprint of life, and cost-effective methods for its long-term storage could have many potential benefits to society. Here we present the method of in situ cryosilicification of whole blood cells, which allows long-term preservation of DNA. Importantly, our straightforward approach is inexpensive, reliable, and yields cryosilicified samples that fulfill the essential criteria for safe, long-term DNA preservation, namely robustness against external stressors, such as radical oxygen species or ultraviolet radiation, and long-term stability in humid conditions at elevated temperatures. Our approach could enable the room temperature storage of genomic information in book-size format for more than one thousand years (thermally equivalent), costing only 0.5 $/person. Additionally, our demonstration of 3D-printed DNA banking artefacts, could potentially allow ‘artificial fossilization’.
研究领域:
生物技术
·
分子生物学
·
基因组学
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ChemistrySelect
·
2022
·Vol. 7
·期 35
·e202202565
·
IF 2.307
Chunying Wei,Haiying Li
Graphical The compounds can bind selectively to G-quadruplex DNA and result in a strong turn-on red fluorescence response, and it is also a potential fluorescence probe for G-quadruplex DNA in the living cells. The small molecule fluorescence probes for G-quadruplexes (G4) play a vital role in the detection of G4. In this work, two novel D−π−A benzothiazole derivatives were synthesized, and their interactions with different secondary structure DNA, cytotoxicity, and intracellular distribution were investigated in detail. Both compounds have a high affinity and about 100-fold turn-on fluorescence response to the promoter G4 in buffer solution, moreover, the compound also stabilizes effectively the promoter G4 structure. Besides, both compounds have no influence on the conformation of the promoter G4, but they induce the folding of bcl-2 from the coiled chain into the mixed-type G4 in salt-free buffer solution. The binding stoichiometries of the compound to bcl-2, c-kit2, and c-myc G4 are near 2 : 1, 1 : 1, and 3 : 1, respectively, and the interactions are an enthalpy-driven process. Furthermore, both compounds inhibit the proliferation of HepG2 cells, with IC 50 values of 3.23 and 38.9 μM. The compound mainly distributes in the cytoplasm of living cells and might interact with DNA. The current results provide the valuable information for design of fluorescence probes targeting G4 in the living cell.
研究领域:
分子生物学
·
药理学
·
生物化学
✓ 已验证
产品关联
DOI
高影响因子
ELECTROCHIMICA ACTA
·
2019
·
IF 5.383
Xia-Xia Liang,Lei Qian,Rong-Fu Huang
Endogenous cell metabolism and chemical reactions can induce oxidative DNA damage, which is involved in the development of different diseases. In this work, a label-free electrochemiluminescence (ECL) assay was proposed for the sensitive detection of oxidative DNA damage in an assembled DNA film. We employed the formamidopyrimidine-DNA glycosylase (Fpg) to convert DNA oxidative nucleobases to strand break, allowing to detect the DNA damage inducing minimal change to the helical structure of DNA. A “light-switch” DNA intercalator, [Ru(bpy) 2 (dppz)] 2+ (bpy = 2, 2 ′ -bipyridine, dppz = dipyrido[3, 2-a:2 ′ , 3 ′ -c]phenazine) was introduced as an ECL signal reporter to detect the DNA strand breaks. The damaged DNA film on the reduced graphene oxide and gold nanoparticles (r-GO@Au) modified ITO electrode bound less ECL indicator than the intact film and accompanied by a drop in ECL intensity, which were also confirmed by gel electrophoresis and fluorescence measurements. As a result, a Fe 2+ -mediated Fenton reaction with as low as 10 nM Fe 2+ and 40 nM H 2 O 2 induced DNA damage can be detected, 100-fold lower than the concentration measured without Fpg. To demonstrate its universality, the successful detection of DNA damage induced by Rose Bengal under irradiation was also achieved. The proposed strategy shows its potential practicality for the sensitive and rapid assessment of the genotoxicity of environmental pollutants .
研究领域:
生物化学
·
分析化学
·
分子生物学
·
环境科学
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2020
·
IF 5.977
Na Wu,Yi-Ting Wang,Xiao-Yan Wang,Xu- Wei Chen,Ting Yang,Jian-Hua Wang
Rapid, sensitive, and user-friendly nucleic acid detection is of growing importance in early clinical diagnosis. Here, we construct a simple, one-pot and ultrasensitive DNA sensor via exonuclease III (Exo III)-assisted target recycling amplification (ERA) combined with 3D DNA walker cascade amplification. In the presence of single-stranded DNA target, the ERA process is activated to generate numerous walker strands (WS). Thereafter, Exo III-powered WSs autonomously move along magnetic bead (MB)-based 3D track to release numerous AgNCs into the supernatant as an amplified signal output. This biosensor had a low detection limit of 18 fM and an analytical range of 40 fM to 1 pM. Furthermore, the practical application potential of this biosensor was also confirmed by the spiking experiments of p53 into human serum and urine samples.
研究领域:
生物技术
·
分子生物学
·
分析化学
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）