INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Yaxue Jia,Siqi Zhao,Anping Wang,Jing Huang,Jinlan Yang,Li Yang
A target-induced multiregion MNAzyme nanowire system is designed for the ultrasensitive and homogeneous detection of microRNAs (miRNAs). miRNA-21 and miRNA-375 are chosen as analytes, and a miRNA-induced primer exchange reaction (PER) is utilized to construct a long DNA strand with repetitive sequences. This innovative design enables the efficient anchoring of numerous MNAzymes. This unique architecture significantly boosts the effective local concentration of MNAzymes, thereby enhancing the sensitivity and efficiency of miRNA detection. Notably, the limit of detection (LOD) achieved with our target-induced multiregion MNAzyme nanowire approach is over an order of magnitude lower than most other MNAzyme-based methods, while the MNAzyme reaction time is reduced from several hours to 50 min. The method has demonstrated successful applications in quantitatively determining the expression levels of two miRNAs in cell lysates of MCF-7, HeLa and MCF-10 A cells, highlighting its potential for assaying miRNA biomarkers in clinical samples.
研究领域:
分子生物学
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生物化学
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诊断学
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ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Feifan Yin,Murilege Chao,Jinhua Chen,Yanheng Yao,Miao He,Qiufeng Wang,Wenting Cheng,Yongliang Cai,Zhongyun Wang,Yang Xiang
mRNA vaccines have been developed as a captivating and highly prospective avenue for the advancement of immunotherapy. With the advancement of antitumor mRNA vaccines into clinical trials, it is important to monitor the levels of nucleic acid drugs. In this study, we have engineered a new biosensor for melanoma-associated antigen mRNA vaccines utilizing an adjustable structure MNAzyme (ASM) activated by an entropy-driven circuit (EDC). The EDC process is specifically triggered by the target mRNA through the exposed toehold, achieving initial amplification. As a result of proximity effects facilitated by the EDC products, ASM is successfully assembled and activated by undergoing conformational transformation because of rational sequence design, continuously cleaving the fluorescence reporter and realizing the second round of signal amplification. Compared with conventional DNAzyme, ASM can effectively reduce signal leakage and concurrently preserve cleavage efficiency, benefiting from the stable presence of a stem-loop hairpin and an adjustable molecular switch. Moreover, composed only of DNA with highly integrated sequences, this method could realize detection in an enzyme-free, one-step, and one-pot detection manner. Finally, EDC-ASM demonstrates high programmability, excellent specificity, and sensitivity in nucleic acid detection, providing a novel strategy for the study of antitumor mRNA vaccines.
研究领域:
分子生物学
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生物传感
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肿瘤免疫治疗
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核酸分析
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转化医学
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药物监测
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TALANTA
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2023
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IF 6.1
Chengjie Duan,Yanheng Yao,Wenting Cheng,Yan Chen,Jin Jiao,Yang Xiang
Aptazyme is a chimera of functional nucleic acids , which integrates recognition and amplification elements to simplify the assay process and improve sensing efficiency. However, its application may be limited by signal leakage. In this work, we innovatively integrate the AβO aptamer and an MNAzyme (multicomponent nucleic acid enzyme) for highly efficient detection of AβO. The aptamer and half of the MNAzyme are positioned at one strand, and the other half of the MNAzyme is integrated with a toehold sequence. These two sequences cannot hybridize to activate the MNAzyme until the target is added. The background signal is significantly reduced by the split format and the secondary structure of DNA probes formed in the absence of the target. The proposed aptazyme can not only achieve amplification through enzymatic catalysis but also greatly improve the efficiency of signal transduction and output. We systematically investigated the influence of different DNA probes on the detection performance, and the optimized aptazyme can detect as low as 26.5 pM targets in 1h. The stability of this method was also investigated by detection targets in real biological samples.
研究领域:
生物化学
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分子生物学
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神经科学
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ANALYTICAL CHEMISTRY
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2026
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IF 7.3
Jiangyan Zhang,Jiajia Li,Yan Wang,Zhaodi Jing,Zhiyong Cheng,Jinding Liu,Hongyuan Yan,Zhengping Li,Yongqiang Cheng
Here, we present a modular and programmable real-time quantitative detection platform that integrates ligase chain reaction (LCR) with multicomponent nucleic acid enzymes (MNAzymes) for single-nucleotide variant (SNV) analysis. By incorporating a universal MNAzyme-based reporter, the LCR–MNAzyme system enables PCR-like real-time quantitative monitoring of SNVs while supporting programmable and multiplex detection. The target recognition module allows precise quantification of multiple variants, while reconfiguring the signal output module enables multiplex detection in a single homogeneous reaction. To further enhance analytical performance, glyoxal-caged probes were incorporated to achieve hot-start LCR, effectively suppressing background amplification and improving detection sensitivity. The optimized system exhibits a linear range from 10 aM to 10 pM with a detection limit of 5.79 aM and enables quantification of the JAK2 V617F mutation down to a variant allele frequency of 0.03%. Quantitative results from clinical samples showed excellent agreement with sequencing data. This work establishes a generalizable framework for highly sensitive, specific, and multiplex SNV detection, advancing the development of quantitative nucleic acid analysis tools for precision molecular research.
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Scientific Reports
·
2026
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IF 4.9
Wang Yingnan,Wang Chang,Zhai Lin,Zhu Xiaoye
Cervical cancer is significantly associated with persistent infection of human papillomavirus (HPV) type 18, making accurate detection of low-abundance HPV-18 mRNA crucial for diagnosis and treatment. Current nucleic acid detection methods are often limited by complex instrumentation, time-consuming procedures, or fluorophore modifications. To address this, we developed an enzyme-free colorimetric biosensor by integrating a multicomponent nucleic acid enzyme (MNAzyme) with catalytic hairpin assembly (CHA) for signal amplification. In this strategy, target recognition triggers the formation of an active MNAzyme that cleaves a substrate hairpin, initiating a CHA cascade which releases G-rich sequences. These sequences form G-quadruplexes with peroxidase-like activity, catalyzing the oxidation of colorless TMB into a soluble blue product, enabling both visual detection and spectrophotometric quantification at 650 nm. Under optimized conditions, the biosensor achieved a detection limit of 6.92 pM for HPV-18 mRNA, allowing naked-eye identification of target levels as low as 10 pM. The platform exhibited high specificity and could be adapted to detect other HPV mRNA subtypes by simply modifying the recognition sequence. This work presents a universal, simplified, and sensitive colorimetric strategy with great promise for future development of point-of-care testing and viral analysis platforms.
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MICROCHIMICA ACTA
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2025
·Vol. 192
·期 3
·1-15
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IF 5.3
Hu Yue,Xia Mengshi,Li Mimi,Li Lulu,Li Chenghong,Liu Yi,Wang Lina,Huang Hui,Fang Lichao,Peng Kexing,Liu Huamin,Wang Xiaolong,Zheng Junsong
The preparation of UiO-66@CdTe@AuNPs composites is presented for the first time, which function as a photoelectrochemical (PEC) sensing matrix and are conjugated to a three-way junction (TWJ). We propose an antibody-based specific recognition-induced neighbor-joining reaction that initiates the assembly of two molecularly designed nucleic acid enzymes (MNAzymes), to release an oligonucleotide that hydrolyzes TWJ through the mechanism of a toehold-mediated strand displacement reaction (TSDR). Subsequently, a hybridization chain reaction application (HRCA)-based dendrimer is formed, which immobilizes a large number of quantum dots to generate a burst effect that reduces the photocurrent signal. As anticipated, the developed PEC biosensor showed excellent analytical performance for both m 6 A-RNA and m 5 C-RNA, with detection limits of 0.309 fM and 6.918 aM, respectively. The successful fabrication of this ultrasensitive and simultaneous PEC biosensor provides new insights for epigenetic research and the bioassay, mechanism investigation and clinical diagnosis of diseases associated with RNA methylation. Graphical
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
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BIOSENSORS & BIOELECTRONICS
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2021
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IF 10.618
Wan Huang,Danyan Zhan,Yiming Xie,Xin Li,Guosong Lai
DNA walkers have been recognized as a type of powerful signal amplification tool for biosensors, but how to adopt a proper strategy to increase their amplification efficiency is still highly desirable. Herein we design a dual-catalytic hairpin assembly (CHA)-mediated strategy for the high-efficient formation of a tripedal Mg 2+ -dependent DNAzyme (MNAzyme)-DNA walker, and thus develop a novel proteinase-free dual-mode biosensing method for the kanamycin (Kana) antibiotic assay. The first CHA is initiated by a target-biorecognition reaction, which can produce the DNA walker and also induce the target recycling. The second CHA is initiated by a special base sequence designed as a one-half substrate of the MNAzyme. Upon the first CHA-triggered DNA walking at a magnetic bead (MB) track, this “pseudo-target” sequence can be released to induce another CHA-cycle for the formation of the same DNA walker. Meanwhile, the other one-half substrate strand exposed on the MB surface will trigger the quantitative hybridization chain reaction (HCR)-assembly of a G-quadruplex DNAzyme (G-DNAzyme)-enriched double-stranded DNA polymer. So the enzymatic reaction of G-DNAzymes enabled the convenient colorimetric and photoelectrochemical dual-mode signal transduction of the method. Due to the dual-CHA facilitation to the tripedal and three-dimensional DNA walking and synergetic signal amplification of HCR, this method exhibits very low detection limits of 9.4 and 0.55 fg mL −1 , respectively. In combination with its wide linear range, automated manipulation, and excellent selectivity , repeatability and reliability, the proposed method is expected to be used for the convenient semiquantitative screening and accurate determination of possible antibiotic residues in complicated matrices.
研究领域:
生物化学
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分析化学
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生物传感器
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纳米技术
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高影响因子
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