BIOSENSORS & BIOELECTRONICS
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2024
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IF 10.7
Chuanyu Yang,Caiyi Du,Feiyu Yuan,Peitong Yu,Boxu Wang,Changshun Su,Ruiqi Zou,Junyang Wang,Xu Yan,Chunyan Sun,Hongxia Li
Benefiting from specific target recognition and trans-cleavage capabilities, the CRISPR/Cas12a system has great application prospects in the design of highly sensitive and rapid fluorescence biosensors. The CRISPR/Cas12a-based fluorophore-quencher molecular beacons exhibit single-color emission and are easily exposed to interference from environmental factors. Herein, we design a CRISPR/Cas12a-derived ratiometric fluorescence sensor for Pb 2+ detection based on embedded carbon dots@zeolitic imidazolate framework-8 (CDs@ZIF-8) composites and DNAzyme. The functions of ZIF-8 about encapsulating red emissive CDs in the inner cavity and adsorbing DNA on the outer surface are integrated to establish dual fluorescence signals, thereby reducing the possibility of interference and improving sensing accuracy. The presence of Pb 2+ is converted into the change of activator by the GR5 DNAzyme to activate the CRISPR/Cas12a system, which provides signal amplification through multiple turnovers of side branch cutting, achieving highly sensitive detection of Pb 2+ with a low detection limit of 18 pM. This method has the advantages of simplicity, universality, and excellent quantitative ability, and has broad prospects in sensing applications.
研究领域:
生物传感
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分子生物学
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环境科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Luyu Wei,Zhilong Wang,Yongzhen Dong,Deyang Yu,Yiping Chen
CRISPR/Cas12a fluorimetry has been extensively developed in the biosensing arena, on account of its high selectivity, simplicity, and rapidness. However, typical CRISPR/Cas12a fluorimetry suffers from low sensitivity due to the limited trans-cleavage efficiency of Cas12a, necessitating the integration of other preamplification techniques. Herein, we develop an enhanced CRISPR/Cas12a fluorimetry via a DNAzyme-embedded framework nucleic acid (FNAzyme) substrate, which was designed by embedding four CLICK-17 DNAzymes into a rigid tetrahedral scaffold. FNAzyme can not only enhance the trans-cleavage efficiency of CRISPR/Cas12a by facilitating the exposure of trans-substrate to Cas12a but also result in an exceptionally high signal-to-noise ratio by mediating enzymatic click reaction. Combined with a functional nucleic acid recognition module, this method can profile methicillin-resistant Staphylococcus aureus as low as 18 CFU/mL, whose sensitivity is approximately 54-fold higher than that of TaqMan probe-mediated CRISPR/Cas12a fluorimetry. Meanwhile, the method exhibited satisfactory recoveries in food matrices ranging from 80% to 101%. The DNA extraction- and preamplification-free detection format as well as the potent detection performance highlight its tremendous potential as a next-generation analysis tool.
研究领域:
生物传感
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分子诊断
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合成生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Yi Deng,Shuqi Quan,Erkang Tian,Jianwen Dong,Meihong Guo,Juan Li,Hao Yang,Shuangquan Lai
Assessment of bacterial viability is vital for food and clinical biosafety, as it reveals pathogen survival and metabolic state, helping predict their potential to persist and cause illness. Herein, we presented a visible light-driven photoelectrochemical assay (termed DzPEC) designed for signal-on detection of live pathogenic bacteria. The DzPEC assay was fabricated using a DNAzyme-functionalized Z-scheme g-C 3 N 4 /V 2 C bio-heterojunction as a photoresposive material and SiO 2 as a photoquencher. The DzPEC assay, which utilizes DNAzyme to target the metabolic endoprotein RNase H2 that is secreted into the extracellular matrix, allows for bacterial viability assessment. Leveraging the Z-scheme heterojunction with high photoactive performance, the DzPEC assay for Salmonella enterica ( S. enterica ), used as a bacteria model, exhibited a detection limit of 141 CFU/mL and the ability to detect live bacterial abundnace as low as 0.1 %. The DzPEC assay demonstrated a strong correlation with RT-qPCR in the detection of S. enterica contamination in complex food and clinical matrices. These findings highlighted the potential of the DzPEC assay in bacterial viability phenotyping and live bacteria-associated biosafety monitoring.
研究领域:
生物传感
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微生物学
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材料科学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Mengke Liu,Peng Li,Haitang Yang,Nan Jiang,Dingzhong Wang,Shihao Sun,Wei Wei
5-hydroxymethylfurfural (HMF) is one of the food contaminants mainly formed by Maillard and caramelization reaction. Here, we developed a simple method to detect HMF based on split-DNAzyme assisted signal amplification via Quartz crystal microbalance . Firstly, two fragments of the split-DNAzyme (ESΙ and ESII) and AuNPs-ssDNA were prepared. In the absence of HMF, the amino group of ESΙ reacted with the aldehyde group of ESΙΙ by Schiff base condensation，which linked ESΙ and ESII as DNAzyme. With the help of Mg 2+ , this DNAzyme cleaved AuNPs-ssDNA, freeing the complementary sequence of capture DNA from the ssDNA. In the presence of the target HMF, the aldehyde group in its molecular structure preferred to react with ESΙ, which prevented the link of ESΙ and ESII. In this case, DNAzyme cannot be formed effectively and the ssDNA modified on AuNPs cannot be cleaved. Secondly, the capture DNA was immobilized on Au chip in the QCM chamber. Then, the intact AuNPs-ssDNA was introduced to hybridize with the capture DNA, resulted in a large frequency change (ΔF) of the QCM biosensor because of the huge increased mass of AuNPs. So, HMF can be detected based on the frequency change (ΔF) of the QCM biosensor. This QCM biosensor is more sensitive than previously reported methods, which provide a new direction for the analytical detection of HMF and other food contaminants.
研究领域:
食品科学
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生物化学
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产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
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2017
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IF 4.95
Chenguang Wang,Nan Cheng,Longjiao Zhu,Yuancong Xu,Kunlun Huang,Pengyu Zhu,Shuifang Zhu,Wei Fu,Wentao Xu
A colorimetric biosensor for DNA screening was designed based on the conformational changes of the horseradish peroxidase (HRP)-mimicking DNAzyme. The scheme of DNA biosensing was designed based on the base pairing of DNAzyme sequence to inhibit the formation of HRP-mimicking hemin/G-quadruplex structures in the process of amplification. DNA could be amplified via the universal primer multiplex polymerase chain reaction (UP- M -PCR) and innovatively detected as color disappear in the reaction visible to the naked eye. The input of key factors and the output of optical characteristics in the reaction inspired the development of an OR logic gate operation for DNA detection. This biosensor overcomes self-inhibition and amplification disparity with the help of UP- M -PCR, thereby exhibiting high specificity and high-throughput without the requirement of gel analysis work. This biosensing system also presented 1% sensitivity and approximately 180 copy numbers in triplicate. The biosensor was used to screen elements from genetically modified organisms (GMOs) and covered more than 90% of all globally authorized events in the world. The designed colorimetric biosensor is a rapid, portable and versatile tool for nucleic acids detection and diagnosis in the field.
研究领域:
生物化学
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分子生物学
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分析化学
✓ 已验证
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2025
·
IF 7.7
Ming Wang,Lizhen Zhao,Yuling Hu,Gongke Li
Shellfish toxins are common marine toxins that have attracted worldwide attention due to their serious threats to food safety and human health. In this work, DNA hydrogel based on rolling circle amplification (RCA) and click reaction was developed for surface enhanced Raman scattering (SERS) detection of okadaic acid (OA) combined with target-responsive DNAzyme signal amplification. The RCA strands containing azido-dCTP were combined with cross-linked strands with ethynyl groups via click reaction to form a DNA hydrogel framework. The click reaction strengthened the DNA hydrogel network, making it more rigid and easier to manipulate during the detection process. The aptamer and DNAzyme were introduced into the DNA hydrogel as functionalized DNA components. Upon the specific recognition of OA by the aptamer, the DNAzyme released via the displacement reaction would continuously cleave the enzyme cutting site within the substrate strand to liberate the SERS signal tag. The functionalized DNA hydrogel concurrently achieved the functions of target recognition, signal conversion and signal amplification. Under optimal conditions, this strategy could detect OA with limits of detection as low as 0.2 nmol/L within 30 min. The SERS signal intensity showed good linear relationship with OA concentration in the range of 1.0–300.0 nmol/L. The practical application was validated for detecting trace amounts of OA in aquatic product samples. The stable DNA hydrogel combined with target-responsive DNAzyme signal amplification offers a universal strategy for rapid detection of trace toxins in complex matrixes.
研究领域:
生物化学
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分析化学
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毒理学
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产品关联
DOI
高影响因子
FOOD CHEMISTRY
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2023
·
IF 8.8
Junjun Wang,Yue Tang,Jia Zheng,Zhengmin Xie,Jianli Zhou,Yuangen Wu
Herein, a colorimetric biosensor for histamine detection in meat is first established based on the enhancement of DNAzyme with peroxidase-mimic activity. Histamine can boost the generation of G-quadruplex sequences, and make them more easily bond with hemin to produce many DNAzyme molecules. In addition, histamine increases the affinity of DNAzyme to the substrate 3,3′,5,5′-tetramethylbenzidine (TMB). Therefore, the obtained DNAzyme can catalyze H 2 O 2 and dissolved oxygen to produce many reactive oxygen species (ROS), which cause the TMB molecule to lose two electrons and generate yellow products, exhibiting a clear absorption peak at 450 nm. The colorimetric biosensor has excellent sensitivity, and the detection limit is as low as 38 μg·L −1 for histamine. Moreover, the biosensor has high selectivity and anti-interference ability, and exhibits a good recovery rate in actual meats. The above results show that the strategy has potential for application in the detection of trace histamine in meats.
研究领域:
生物化学
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食品科学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Yanju Chen,Hui Wu,Siwenjie Qian,Xiaoping Yu,Huan Chen,Jian Wu
Heavy metal lead accumulation in the environment pollutes the ecology systems and further threatens the human health. It is necessary to develop a sensitive method to detect it. Here, we propose a highly sensitive lead detection method by combining DNAzyme and CRISPR system. Once the lead ion is recognized, the substrate chain of DNAzyme is cleaved to produce single strand DNA . The produced single strand DNA can be detected by Cas protein/guide RNA complex and further trigger the collateral cleavage effect of CRISPR system, which can indiscriminately cut short single strand DNA reporters. By this way, the detection signals can be greatly amplified. This method can detect lead ions as low as 0.48 nM. The sensitivity is higher than the DNAzyme method. Furthermore, the portable 3D printing device is designed to observe the fluorescent signals so the end-point detection results can be visualized by the naked eyes. The entire detection process can avoid using bulky and expensive instruments, which can promote on-site lead ion detection.
研究领域:
生物化学
·
分子生物学
·
环境科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Li Zhang,Zhang Zhang,Xinying Ou,Yufeng Wang,Liu Yang,Zhi Weng,Guoming Xie
Single nucleotide variants (SNVs) are essential biomarkers both clinically and biologically, and the specific hybridization of matched sequences is crucial to discriminate SNVs. However, the similar sequence would hybridize to probes incorrectly and cause false positive. Herein, we designed a DNAzyme-based nanomachine guided by thermodynamic and kinetic parameters to identify SNVs. The standard Gibbs free energy of the internal loop in the blocked DNAzyme was rationally taken into consideration and a quasi-steady 'toehold exchange' reaction was designed. These simulations in silico enable an optimal performance to discriminate SNVs for various conditions. Additionally, we introduced a DNA fuel to enable non-covalent DNA catalysis reaction, which can tune the performance of nanomachine by changing the concentration of DNA fuel. In this way, we make a satisfactory trade-off between sensitivity and specificity to discriminate SNVs. Collectively, this nanomachine produced discrimination factors between 5 and 31, the detection limit of 0.1 % variant alleles can also be identified. Our work demonstrated a pathway assist to the design of DNA nanomachine through simulation in silico, which hold great potential for the application of complex DNA nanomachines and biosensors.
研究领域:
生物化学
·
分子生物学
·
纳米技术
✓ 已验证
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2022
·
IF 5.895
Chuanyu Yang,Peitong Yu,Ying Li,Junyang Wang,Xinyue Ma,Ni Liu,Ting Lv,Hongru Zheng,Han Wu,Hongxia Li,Chunyan Sun
Lead contamination has posed a potential threat to the environment and food safety, arousing extensive concern. In this work, we fabricated a novel fluorescent sensing platform based on zeolitic imidazolate framework-8 (ZIF-8) and DNAzyme for monitoring Pb2+ in water and fish samples. ZIF-8 was proposed as a fluorescence quencher with the advantages of simple synthesis, low cost, and high quenching efficiency. The Pb2+-dependent GR5 DNAzyme containing the large ssDNA loop can be adsorbed onto ZIF-8 accompanied by fluorescence quenching. Upon binding with Pb2+, GR5 DNAzyme was activated and cleaved, leading to the release of FAM-labeled 5-base ssDNA, which restored the fluorescence. The “turn-on” assay can detect Pb2+ through the one-pot procedure in the range of 0.01–10.0 nM with a detection limit of 7.1 pM. The platform is promising for on-site monitoring of Pb2+ owing to the excellent performance of high sensitivity, low background, strong anti-interference ability, and simple operation.
研究领域:
分析化学
·
环境科学
·
生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Acta Biomaterialia
·
2023
·
IF 9.7
Jiawu Yu,Yuchen Zhang,Liqi Li,Yang Xiang,Xuemei Yao,Youbo Zhao,Kaiyong Cai,Menghuan Li,Zhongjun Li,Zhong Luo
Radiotherapy is a mainstream modality for breast cancer treatment that employs ionizing radiation (IR) to damage tumor cell DNA and elevate ROS stress, which demonstrates multiple clinically-favorable advantages including localized treatment and low invasiveness. However, breast cancer cells may activate the p53-mediated cell cycle regulation in response to radiotherapy to repair IR-induced cellular damage and facilitate post-treatment survival. F-Box and WD Repeat Domain Containing 7 (FBXW7) is a promoter of p53 degradation and critical nexus of cell proliferation and survival events. Herein, we engineered a cooperative radio-ferroptosis-stimulatory nanomedicine through coordination-driven self-assembly between ferroptosis-inducing Fe 2+ ions and FBXW7-inhibiting DNAzymes and further modification of tumor-targeting dopamine-modified hyaluronic acid (HA). The nanoassembly could be selectively internalized by breast cancer cells and disintegrated in lysosomes to release the therapeutic payload. DNAzyme readily abolishes FBXW7 expression and stabilizes phosphorylated p53 to cause irreversible G2 phase arrest for amplifying post-IR tumor cell apoptosis. Meanwhile, the p53 stabilization also inhibits the SLC7A11-cystine-GSH axis, which combines with the IR-upregulated ROS levels to amplify Fe 2+ -mediated ferroptotic damage. The DNAzyme-Fe-HA nanoassembly could thus systematically boost the tumor cell damaging effects of IR, presenting a simple and effective approach to augment the response of breast cancer to radiotherapy. Statement of Significance To overcome the intrinsic radioresistance in breast cancer, we prepared co-assembly of Fe 2+ and FBXW7-targeted DNAzymes and modified surface with dopamine conjugated hyaluronic acid (HA), which enabled tumor-specific FBXW7-targeted gene therapy and ferroptosis therapy in IR-treated breast cancers. The nanoassembly could be activated in acidic condition to release the therapeutic contents. Specifically, the DNAzymes could selectively degrade FBXW7 mRNA in breast cancer cells to simultaneously induce accumulation of p53 and retardation of NHEJ repair , eventually inducing irreversible cell cycle arrest to promote apoptosis. The p53 stabilization would also inhibit the SLC7A11/GSH/GPX4 axis to enhance Fe 2+ mediated ferroptosis. These merits could act in a cooperative manner to induce pronounced tumor inhibitory effect, offering new approaches for tumor radiosensitization in the clinics.
研究领域:
肿瘤学
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分子生物学
·
纳米医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2023
·
IF 6.1
Ying Liao,Yingqin Zhang,Aiwen Su,Yanli Zhang,Hongbin Wang,Wenrong Yang,Pengfei Pang
Protein kinase A (PKA) can regulate many cellular biological processes by phosphorylation substrate peptide or protein. Sensitive detection of PKA activity is critical for the PKA-related drug discovery and disease diagnosis. A new electrochemical biosensing method was developed for detection of PKA activity based on Zr 4+ -mediated DNAzyme-driven DNA walker signal amplification strategy. In this strategy, the special designed substrate peptide and a thiolated methylene blue-labeled hairpin DNA (MB-hpDNA) containing a single ribonucleic acid group (rA) could be anchored on the surface of gold electrode by Au–S bond. In the presence of adenosine triphosphate (ATP) and PKA, substrate peptide was phosphorylated and linked with walker DNA (WD) via the robust phosphate-Zr 4+ -phosphate chemistry. The linked WD hybridized with the loop region of MB-hpDNA to form a Mn 2+ -dependent deoxynuclease (DNAzyme), which cleaved the MB-hpDNA into MB-labeled fragment releasing away from electrode surface, resulting in a dramatic decrease of electrochemical signal and providing an electrochemical sensing platform for PKA activity detection. The response signal of the developed biosensor is proportional to the logarithm of PKA concentration in the range of 0.05 U mL −1 to 100 U mL −1 , with a detection limit of 0.017 U mL −1 at a signal to noise ratio of 3. Furthermore, the proposed method can also be applied for the evaluation of PKA inhibition and PKA activity assay in cell samples. Therefore, the proposed biosensor shows great promise as a universal tool for diagnostics and drug discovery of PKA-related diseases.
研究领域:
生物化学
·
分析化学
·
医学诊断
✓ 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）