TALANTA
·
2025
·
IF 6.1
Xiang Li,Yuting Zhang,Mengyuan He,Jiarui Sun,Bowen Xiong,Gongke Wang
In this work, we present an ultrasensitive, specific, and high-signal-to-background ratio fluorescence split-aptasensor for D-vasopressin (D-VP) detection. This sensor is based on target-induced self-propelled 3D DNA walkers in conjunction with CRISPR-Cas12a technology. Two split probes (SDA 1 and SDA 2) were designed to undergo structural recombination and function as a walking chain (SDA) under the induction of D-VP. Simultaneously, an intact Mg 2+ -dependent DNAzyme domain was formed at the tail of SDA and subsequently activated. The activated Mg 2+ -dependent DNAzyme continuously propelled the 3D DNA walker, enabling the generation of signal strand DNA (activator DNA). The activator DNA can subsequently trigger the activation of the Cas12a protein, enabling it to cleave the FAM-ssDNA-BHQ1 substrate. This process leads to signal amplification and the specific detection of D-VP. Under optimal conditions, the designed split-aptasensor exhibits excellent linearity across a concentration range of 5 ng/mL to 1215 ng/mL, with a detection limit (LOD) as low as 0.22 ng/mL. This split-aptasensor were employed to identify D-VP in human serum and urine samples, yielding highly satisfactory results. This unique design acts as a proof of concept and illustrates considerable promise for the detection of a wide range of analytes.
研究领域:
生物化学
·
分子诊断
·
纳米生物技术
✓ 已验证
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Yunshan Zhang,Qianglong Tan,Fang Yang,Tuo Huang,Siyu Yu,Jing Ye,Jianxian Zeng,Xianzhong Feng,Diming Zhang
DNA walkers, as structurally and functionally programmable signal amplification tools, exhibit great potential for application in the field of biosensing. Traditional DNA walkers often rely on enzymes for operation, posing compatibility challenges, while the handful of existing enzyme-free DNA walkers demonstrate limited performance. To address this, we innovatively developed an efficient enzyme-free 3D DNA walker with dual capture and dual output capabilities. Coupled with ligase chain reaction (LCR), this system facilitates highly sensitive and specific detection of single nucleotide polymorphisms (SNPs). Specifically, LCR precisely identifies single-base mutations, effectively transmitting biological information. The 3D DNA walker system is based on entropy-driven circuit cycling reaction technology. In this system, LCR products serve as the driving strands for the DNA walker, independently binding to track strands and walking legs immobilized on gold nanoparticles, forming a unique dual signal capture mechanism. Each track strand carries two signal chains, significantly enhancing signal amplification efficiency. Benefiting from this novel enzyme-free 3D DNA walker strategy, our biosensing system exhibits exceptional sensitivity to mutant targets (MT), detecting MT at concentrations as low as 30.3 aM and distinguishing heterozygous samples with a 0.01% mutation frequency. Furthermore, this system has been successfully applied to genotyping and mutation abundance assessment of genomes from fresh soybean leaves, demonstrating its vast potential for practical applications. In summary, this research pioneers a novel enzyme-free 3D DNA walker with dual capture and dual output capabilities, and develops an ultrasensitive genotyping tool. This provides strong technical support for the advancement of genetic research.
研究领域:
生物传感
·
遗传学
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Linjing Duan,Huali Jin,Yang Wen,Min Wei,Rui Guo,Zhiguang Suo,Keshavan Niranjan
As a food and environmental contaminant having harmful qualities like neurotoxicity and carcinogenicity, acrylamide (AA) is a risk to human health. In this study, Ce-MOF@AuPd NFs were utilized as electrode modification materials to develop a signal-off electrochemical aptasensor in combination with Mg 2 + -dependent DNAzyme-driven 3D DNA walker and triplex molecular switch (THMS) for the sensitive detection of AA. Ce-MOF@AuPd NFs exhibit an expansive surface area and enhanced conductive properties, enabling the binding of substantial DNA sequences and speeding up electron transfer. The Mg 2+ -dependent DNAzyme-driven 3D DNA walker improves the nucleic acid shearing efficiency with excellent signal amplification performance, and combines with the DNA-specific structure THMS to further boost sensor sensitivity. The sensor also demonstrated excellent specificity, reproducibility, repeatability, and stability, along with achieving satisfactory recovery rates when tested on potato chips and coffee samples. The electrochemical aptasensor offers strong application prospects for AA detection in food, according to the results.
研究领域:
生物传感技术
·
食品安全检测
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2022
·
IF 14.224
Shuo Qi,Xiaoze Dong,Yuhan Sun,Yin Zhang,Nuo Duan,Zhouping Wang
DNA walker machines, as one of the dynamic DNA nanodevices, have attracted extensive interest in the field of analysis due to their inherent superiority. Herein, we reported a split aptamer remodeling-initiated target-self-service 3D-DNA walker for ultrasensitive, specific, and high-signal-background ratio determination of 17β-estradiol (E2) in food samples. Two split probes (STWS-a and STWS-b) were rationally designed that can undergo structural reassembled to serve as walking strands (STWS) under the induction of the target. Meanwhile, an intact E6-DNAzyme region was formed and activated at the tail of STWS. The activated E6-DNAzyme then continuously drives the 3D-DNA walker for signal amplification and specific detection of E2. Under optimal conditions, the proposed DNA walker-based biosensor exhibited excellent linearity in the range of 1 pM to 50 nM with a low limit of detection (LOD) of 0.28 pM, and good precision (2.7%) for 11 replicate determinations of 1 nM of E2. Furthermore, the developed DNA walker-based biosensor achieved excellent sensitive analysis of E2 in the complex food matrix with recoveries of 95.6–106.5%. This newly proposed split aptamer-based strategy has the advantages of ultrasensitive, high signal-to-background ratio, and high stability. Noteworthy, the successful operation of the DNA walker initiated by the split aptamer expands the principles of DNA walker design and provides a universal signal amplification platform for trace analysis.
研究领域:
分析化学
·
纳米技术
·
生物传感器
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Kai Zhang,Zhenqiang Fan,Yue Huang,Yuedi Ding,Minhao Xie
Early diagnosis and timely management of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) are the keys to preventing the spread of the epidemic and controlling new infection clues. Therefore, strengthening the surveillance of the epidemic and timely screening and confirming SARS-CoV-2 infection is the primary task. In this work, we first proposed the idea of activating CRISPR-Cas12a activity using double-stranded DNA amplified by a three-dimensional (3D) DNA walker. We applied it to the design of an electrochemiluminescent (ECL) biosensor to detect the SARS-CoV-2 RNA-dependent RNA polymerase (RdRp) gene. We first activated the cleavage activity of CRISPR-Cas12a by amplifying the target DNA into a segment of double-stranded DNA through the amplification effect of a 3D DNA walker. At the same time, we designed an MXene based ECL material: [email protected] 3 C 2 @AuNPs, and constructed an ECL biosensor to detect the RdRp gene based on this ECL material as a framework. Activated CRISPR-Cas12a cleaves the single-stranded DNA on the surface of this sensor and causes the ferrocene modified at one end of the DNA to move away from the electrode surface, increasing the ECL signal. The extent of the change in electrochemiluminescence reflects the concentration of the gene to be measured. Using this system, we detected the SARS-CoV-2 RdRp gene with a detection limit of 12.8 aM. This strategy contributes to the rapid and convenient detection of SARS-CoV-2-associated nucleic acids and promotes the clinical application of ECL biosensors based on CRISPR-Cas12a and novel composite materials.
研究领域:
生物技术
·
纳米技术
·
分子生物学
·
病毒学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 11
·1-11
·
IF 5.3
Wei Yuxin,Fu Yang,Li Chuanhai,Chen Siyu,Xie Linzhi,Chen Mei
A electrochemical biosensor was designed utilizing a CRISPR Cas9n-driven DNA walker combined with gold-nanosphere-like covalent organic frameworks (COFs-AuNPs) to detect breast cancer markers (PIK3CA E545K ctDNA). The DNA walker probe is activated only in the presence of circulating tumor deoxyribonucleic acid (ctDNA), binding to a support probe to form a double strand that is then specifically cleaved by the Cas9n/sgRNA complex. This cleavage produces numerous DNA fragments for signal amplification. The COF-AuNPs as electrode materials facilitate electronic transfer and provide additional active sites for the immobilization of nucleic acid probes. This setup achieves a detection limit of 1.76 aM, demonstrating high sensitivity. Additionally, Cas9n improves the specificity of the sensor, accurately distinguishing a pair of base-mismatched sequences, and reducing the occurrence of false positives. Overall, the sensor exhibits excellent selectivity, reproducibility, and potential for early diagnosis of breast cancer. Graphical Abstract
研究领域:
生物技术
·
癌症生物学
·
纳米技术
·
电化学
·
分子诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Chemistry-An Asian Journal
·
2023
·
IF 4.1
Junyou Li,Hung-Wing Li
By combining exquisitely designed hairpins with the catalytic hairpin assembly to form tripedal DNA walkers driven by enzyme, we constructed a 3D DNA walker with accordingly complementary hairpins attached on gold nanoparticles (AuNPs) and sensitive fluorescence sensing system for the sensitive detection of target miRNA-21. The presence of miR-21 triggers the CHA among three hairpins, which lead to the formation of the tripedal DNA walkers. For the walking trajectories, FAM-labeled hairpins were attached to the surface of AuNPs, the fluorescence of which was initially quenched due to its close proximity to AuNPs. After the binding/cleaving/moving process of tripedal DNA walkers with HP4 driven by Exonuclease III (Exo III), a number of single-stranded DNAs (ssDNAs) will be released with FAM fluorescence recovered. Benefiting from the DNA walker and CHA cascade amplification, the proposed sensing strategy showed remarkable improvement in sensitivity with the LOD of 46 aM. Also, the system was practically applied in human serum samples, demonstrating satisfactory outcome. Owing to the precise design of the system, this method exhibited excellent specificity to distinguish miR-21 from its single-, double-mismatched sequences and non-complementary sequences, showing great versatility and potential for the biological analysis and early disease diagnosis.
研究领域:
生物技术
·
纳米技术
·
癌症诊断
·
分子生物学
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）