MICROCHEMICAL JOURNAL
·
2026
·
IF 5.2
Feifei Zhou,Yingzhan Fan,Qinrui Cao,Yingying Yuan,Yingxiao Chen,Lingling Liu,Jingwen Mai,Nandi Chen,Tianxing Ji,Dou Wang,Chenchen Ge
Dual-function DNA probe (dfDNA) enables a split crRNA/Cas12a (SCas12a) assay. • The dfDNA acts as both a Cas12a activator and a fluorescent reporter. • The dfDNA activates both the cis - and trans -cleavage activities of Cas12a. • The dfDNA/SCas12a assay detects microRNA-21 (miR-21) without amplification. • Bead-assisted dfDNA format improves the sensitivity for miR-21 detection. MicroRNA-21 (miR-21) is a promising non-invasive biomarker for tumor diagnosis, but its short length and low abundance make amplification-free detection challenging. Here, we developed a dual-function DNA probe (dfDNA)-enabled split crRNA-Cas12a (SCas12a) system for miR-21 detection. This design separates crRNA into a constant scaffold fragment (sc-crRNA) and a variable spacer fragment (sp-crRNA), and miR-21 functionally replaces the spacer fragment to activate Cas12a in the presence of dfDNA. The dfDNA acts as both a target-recognition probe and a fluorescent reporter: it hybridizes with miR-21, activates Cas12a, and presents a single-stranded fluorescent overhang that can be cleaved after Cas12a activation. Because target recognition and signal reporting are integrated within a single probe, dfDNA positions the activated Cas12a complex close to the cleavable overhang, improving signal conversion compared with the conventional single-stranded DNA reporter and fully complementary double-stranded DNA probe format. Under optimized conditions, the limit of detection (LOD) with the homogeneous ssDNA reporter-based assay was 347.9 pM for miR-21, whereas the homogeneous dfDNA-based assay reduced the LOD to 19.8 pM. When transferred to a magnetic bead (MB)-assisted format, the dfDNA-based system achieved an LOD of 1.1 pM, supporting that the localized reaction accelerates the trans -cleavage kinetics of Cas12a. The MB-assisted dfDNA assay achieves comparable or even superior sensitivity for miR-21 detection compared with previously reported split crRNA-based fluorescence methods. Further optimization, including target enrichment or signal amplification strategies, is needed before this method can be reliably used for low-abundance miRNAs in clinical serum samples. Download: Download high-res image (88KB) Download: Download full-size image
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2026
·
IF 11.8
Jingtao Huang,Yu Ting Xu,Jin Chen Liu,Wen Yi Lv,Li Ping Cao,Hong Yan Zou,Chun Mei Li,Cheng Zhi Huang
The 10-23 DNAzyme is a potent catalytic nucleic acid for biosensing; however, traditional single-blocker designs suffer from high background signal leakage due to spontaneous thermal “breathing.” To overcome this, we engineered a triangular DNA probe-DNAzyme (TDPzyme) that spatially separates the enzyme components onto a rigid 3D scaffold. By splitting the binding arms and catalytic center, the TDPzyme functions as a fault-tolerant logic “AND” gate, strictly requiring the simultaneous displacement of multiple blocker strands by the target miR-21. This architectural constraint physically shields the probe from nuclease degradation and virtually eliminates false-positive activation, yielding a near-zero background state. Functionalized with a Cy3-BHQ2 pair and an AS1411 aptamer for targeted, carrier-free cellular delivery, the TDPzyme achieved an approximately 18-fold enhancement in sensitivity over conventional closed-arm designs, establishing a linear range of 0.1 nM to 40.0 nM and a limit of detection of 50.3 pM (3σ). Furthermore, time-lapse dynamic imaging demonstrated that the TDPzyme enables highly reliable, high-contrast discrimination of miR-21 expression between tumor and normal cells. Ultimately, the TDPzyme design offers a robust, structurally controllable platform for developing highly sensitive, low-background biosensors applicable to demanding live-cell imaging and precision molecular diagnostics.
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高影响因子
Journal of Materials Chemistry B
·
2017
·Vol. 5
·期 34
·7133-7139
·
IF 4.543
Yike Fu,Tong Chen,Gang Wang,Tongxu Gu,Congkun Xie,Jie Huang,Xiang Li,Serena Best,Gaorong Han
MicroRNAs (miRNAs) play a key role in regulating gene expression but can be associated with abnormalities linked to carcinogenesis and tumor progression. Hence there is increasing interest in developing methods to detect these non-coding RNA molecules in the human circulation system. Here, a novel FRET miRNA-195 targeting biosensor, based on silica nanofibers incorporated with rare earth-doped calcium fluoride particles (CaF2:Yb,Ho@SiO2) and gold nanoparticles (AuNPs), is reported. The formation of a sandwich structure, as a result of co-hybridization of the target miRNA which is captured by oligonucleotides conjugated at the surface of CaF2:Yb,Ho@SiO2 fibers and AuNPs, brings the nanofibers and AuNPs in close proximity and triggers the FRET effect. The intensity ratio of green to red emission, I541/I650, was found to decrease linearly upon increasing the concentration of the target miRNA and this can be utilized as a standard curve for quantitative determination of miRNA concentration. This assay offers a simple and convenient method for miRNA quantification, with the potential for rapid and early clinical diagnosis of diseases such as breast cancer.
研究领域:
生物传感器
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纳米技术
·
分子生物学
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癌症研究
·
分析化学
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PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Fangming Chen,Lu Yang,Feng Fu
The development of miniaturized, label-free microRNA (miRNA) sensors is of great significance for early cancer diagnosis, disease monitoring, and point-of-care testing (POCT), which can substantially enhance clinical efficiency and diagnostic accuracy. In this work, we proposed a functional nucleic acid-based miniaturized homogeneous electrochemical biosensor for label-free miRNA detection. The system features an integrated microelectrode (IME) comprising a gold fiber microelectrode paired with carbon rod/Ag/AgCl wires within a three-channel glass microtube, enabling electrochemical monitoring at the microliter scale. A functional nucleic acid probe containing a miRNA-complementary DNA sequence and a G-triplex (G3) motif was synthesized. Leveraging the strong affinity between G3 and methylene blue (MB), the G3/MB complex serves as a signal generator by significantly reducing MB diffusion current upon binding. Using microRNA21 (miR21) as the model target, the biosensor achieved a detection limit of 0.78 nM with excellent specificity. Successful detection of miR21 in diluted human serum and MCF-7 cell lysate samples confirmed its applicability in complex matrices. This work establishes a “Mix and Measure” platform for homogeneous, label-free electrochemical miRNA biosensing.
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高影响因子
MICROCHIMICA ACTA
·
2025
·Vol. 192
·期 2
·1-9
·
IF 5.3
Su Yuan,Zhang Ziyu,Zhang Hongfang
A biosensor based on solid-state nanochannels of anodic aluminum oxide (AAO) membrane for both electrochemical and naked-eye detection of microRNA-31 (MiR-31) is proposed. For this purpose, MoS 2 nanosheets, which possess different adsorption capabilities to single-stranded and double-stranded nucleic acids, are deposited onto the top surface of the AAO membrane. Moreover, multi-functional DNA nanostructure have been designed by linking a G-rich sequence for folding to a G- quadruplex at three vertices and a complementary sequence of MiR-31 at the other one vertex of a DNA tetrahedron. In the absence of MiR-31, the tetrahedron DNAzyme probe formed after the addition of hemin can mediate the deposition of insoluble on MoS 2 /AAO, which not only enables the color change of the membrane but also gates the transport of K 3 [Fe(CN) 6 ] across the nanochannels. Therefore, the detection of MiR-31 is realized by both visual observation of the brown color and measuring the electrochemical redox current of [Fe(CN) 6 ] 3 − . Using this biosensor, a detection limit as low as 0.06 fM is achieved. The dual-mode detection method also exhibits good specificity, reproducibility, and stability, demonstrating potential application in the diagnosis of oral squamous cell carcinoma and other related biological purposes. Graphical
研究领域:
生物传感器
·
分子诊断
·
纳米技术
·
癌症研究
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PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2020
·
IF 10.257
Fengyun Li,Gen Li,Shijie Cao,Boshi Liu,Xiaoliang Ren,Ning Kang,Feng Qiu
MicroRNAs (miRNAs) are important biomarkers for the diagnosis, prognosis, and treatment of human diseases. Sensitive and selective detection of multiple miRNAs simultaneously will greatly facilitate the early and accurate diagnosis of cancers. Herein, a novel entropy-driven amplification system-templated silver nanoclusters sensing platform was developed for the multiplexed analysis of tumor-associated miRNAs. The sensing platform was constructed by coupling target-triggered entropy-driven catalysis with luminescence adjustable DNA-templated silver nanoclusters (Ag NCs). In the presence of target miRNA, the sensing platform initiates the branch migration and strand displacement of the complex, which has a six-base cytosine loop for stabilizing the luminous Ag NCs. The target is cyclically generated for new catalysis while turning off the fluorescence of Ag NCs; this is accompanied by a significantly amplified optical readout. In this study, two different complex-stabilized Ag NCs systems were proposed, the yellow-emitting Ag NCs and red-emitting Ag NCs biosensors enabled the analysis of miRNA-141 and miRNA-155 with detection limits of 6.1 pM and 8.7 pM, respectively. Impressively, owing to the excellent selectivity , flexibility, and narrow-band excitation of the platform, the multiplexed synchronous detection of miRNA-141 and miRNA-155 were achieved in buffer, biological cell lysates and human serum samples with satisfactory results. The simple, flexible, and convenient strategy provides a powerful tool for multiple biomarkers analysis and related clinical applications.
研究领域:
生物标志物研究
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纳米技术
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癌症诊断
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分子生物学
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分析化学
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PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2019
·
IF 8.355
Tongxu Gu,Zhengyan Li,Zhaohui Ren,Xiang Li,Gaorong Han
Simultaneous detection of multiple types of microRNA (miRNA) is of great significance in early cancer diagnosis. In this study, rare-earth-doped CaF 2 upconversion nanocrystals (CaF 2 : RE 3+ , namely Ho 3+ or Tm 3+ and Yb 3+ ) are formed within the mesopores of silica nanoparticles (CaF 2 : RE 3+ @MSN), in an in-situ manner, by capillary effect and thermo-decomposition method. Comparing to current upconversion nanoparticles, the amorphous and mesoporous nature of CaF 2 : RE 3+ @MSN provides abundant active groups and high specific surface area for feasibly immobilizing oligonucleotide probe. The simultaneous and selective detection of multiple miRNAs was achieved based on the photoluminescence spectra of different CaF 2 : RE 3+ @MSN conjugated with DNA probes in the same solution. The results indicated that the proposed approach holds significant potential for multiple miRNA detection and offers an opportunity for cancer early diagnosis.
研究领域:
纳米技术
·
生物医学工程
·
分析化学
·
分子生物学
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高影响因子
ANALYST
·
2018
·Vol. 143
·期 15
·3702-3707
·
IF 3.864
Jun-Tao Cao,Fu-Rao Liu,Fang Hou,Juan Peng,Shu-Wei Ren,Yan-Ming Liu
The cathodic electrochemiluminescence (ECL) behaviour of nontoxic MoS2 quantum dots (QDs) was studied for the first time using potassium peroxydisulfate as the co-reactant. Ag-PAMAM NCs, serving as difunctional tags for quenching and enhancing ECL of MoS2-reduced graphene oxide composites, were introduced into the ECL detection system for signal amplification. By modulating the interparticle distance between MoS2 QDs and Ag-PAMAM NCs, the ECL quenching from resonance energy transfer and the ECL enhancement from surface plasma resonance were realized. Coupling the good ECL performance of MoS2 QDs with the excellent ECL quenching and enhancement effects of Ag-PAMAM NCs, a novel MoS2 QDs-based ECL biosensing platform for sensitive detection of microRNA-21 was achieved with a detection limit of 0.20 fmol L−1 (S/N = 3). This method was successfully applied to the determination of microRNA-21 in human serum samples with recoveries of 90.0–110.0%, suggesting great potential for its applications in biological and chemical analysis.
研究领域:
分析化学
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生物化学
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纳米技术
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分子生物学
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高影响因子
JOURNAL OF SOLID STATE ELECTROCHEMISTRY
·
2015
·Vol. 20
·期 2
·413-419
·
IF 2.446
Zhou Yunlei,Wang Mo,Yang Zhiqing,Lu Linan,Yin Huanshun,Ai Shiyun
Nuclease S1 can catalyze the nonspecific endo- and exonucleolytic cleavage of single-stranded DNA and RNA to yield nucleoside 5′-phosphates and 5′-phosphooligonucleotides. However, it cannot hydrolyze double-stranded DNA, double-stranded RNA, or DNA-RNA hybrid. Inspired by this specific property, a simple electrochemical method was developed for microRNA detection based on hybridization protection against nuclease S1 digestion. In the absence of hybridization process, the assembled probe DNA on the electrode surface can be easily digested by nuclease S1 and a strong electrochemical signal can be generated due to the decreased repulsive force towards the redox probe. However, after hybridization with target microRNA, the digestion activity of nuclease S1 is inhibited, which can lead to a weak electrochemical signal. Based on the change of the electrochemical signal, the detection of target microRNA-319a can be achieved. Under optimal experiment conditions, the electrochemical signal was proportional to microRNA-319a concentration from 1000 to 5 pM and the detection limit was 1.8 pM (S/N = 3). The developed method also showed high detection selectivity and reproducibility. Furthermore, the proposed method was successfully applied to assay the expression level of microRNA-319a in the leaves of rice seedlings after being incubated with different concentrations of 6-benzylaminopurine.
研究领域:
生物化学
·
分析化学
·
植物生物学
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高影响因子
JOURNAL OF APPLIED ELECTROCHEMISTRY
·
2022
·Vol. 52
·期 5
·885-893
·
IF 2.925
Zhao Fei,Xue Wenwen,Zhang Hongfang
Abnormal expression and functioning of microRNAs in the immune system are frequently linked to various cancers. Our previous work demonstrated a universal, label-free, and reliable microRNA detection platform based on the nanochannels-based recognition interface and electrode-based sensing interface. Herein, the strategy of G-quadruplex DNAzyme-catalyzed deposition was introduced into the nanochannels to improve the detection sensitivity of the platform. G-quadruplex DNAzyme could catalyze the oxidation of 4-chloro-1-naphthol to produce insoluble precipitates into the nanochannels which amplified the effect of the hybridization reaction between the capture probe and the target to the mass transport of the redox probe through the recognition interface. Then, a porous carbon nanofibers-modified electrode was applied to further amplify the monitoring sensitivity of the sensing interface to the concentration change of the redox probe methylene blue. Thereby, sensitive electrochemical detection of microRNA-21 was acquired. The biosensing platform exhibited a broad linear range of 100 aM to 1 nM for microRNA-21, with a low detection limit of 40 aM. What is more, the selective and reproducible platform provided an alternative technique for the PCR-free determination of microRNA in serum and tumor cells. Graphical abstract
研究领域:
生物化学
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分子生物学
·
癌症研究
·
电化学
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 20
·6157-6166
·
IF 4.478
Liang Yuling,Yang Huihui,Yin Wen,Zhang Yanfei,Xu Yuzhi,Liu Si-Yang,Dai Zong,Zou Xiaoyong
Long-term and continuous monitoring of the microRNA (miRNA) expression in living cells is essential in biomedical research, but it is currently limited by fast consumption and easy digestion of probes in the intracellular environment. Herein, we report polydopamine-modified gold nanoparticles (AuNPs@PDA) as protective and efficient nanocarriers for DNA hairpin probes (hpDNA), achieving long-term monitoring (48 h) of the miRNA (let-7a) levels in living cells after drug treatments. This method enabled excellent sensitivity and high selectivity toward let-7a with a limit of detection of 0.51 nM ( n = 3) and a linear range from 1 to 100 nM. More importantly, AuNPs@PDA can not only efficiently improve the loading of hpDNA on each nanoparticle, but also effectively protect hpDNA from hydrolysis in the cell microenvironment, finally realizing the continuous monitoring of let-7a in living cells for 48 h. This simple method would be of great significance for drug screening and precision medicine. Graphical abstract
研究领域:
生物医学工程
·
纳米技术
·
分子生物学
·
药物发现
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PubMed
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2022
·Vol. 32
·期 48
·2207879
·
IF 19.924
Lexiang Zhang,Wenzhao Li,Rokshana Parvin,XiaoChen Wang,Qihui Fan,Fangfu Ye
Exosomes are cell-derived extracellular vesicles which enclose genetic substances from their origin cells, offering a good source of biomarkers. Many efforts have been made to develop non-invasive exosome-based clinical diagnosis, which are urgently needed for prostate cancer, as traditional methods are criticized due to the inaccuracy and discomfort. In this study, a simple, universal process for genotyping of exosome-derived microRNAs using nanoparticles to capture exosomes and the novel photothermal droplet digital polymerase chain reaction (ddPCR) assay to detect target molecules is presented. To detect the target sensitively and simply, the smart ddPCR thermocycling of the agarose microcarriers can be controllably realized via embedding photothermal MXene and magnetic nanoparticles composite, and then irradiating with a homemade automated near-infrared control module. Duplex photothermal-driven ddPCR cooperating with dual TaqMan probes enabled simultaneously and quantitatively detecting copy numbers of prostate oncogenes microRNA-21-5p, microRNA-375-3p, and metastasis microRNA-574-3p along with the control microRNA-30a-5p within the extracted exosomes. The results show that the expression levels of these biomarkers in prostate cancer cell lines PC-3 and DU145 are 7–210 folds higher than those in healthy control cells HaCaT and HUVEC. Thus, it is believed that these versatile exosomal microRNA genotyping approaches are potentially valuable for diverse clinical applications.
研究领域:
癌症生物学
·
分子诊断
·
纳米技术
·
生物医学工程
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高影响因子
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