APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2024
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IF 3.1
Yan Na,Hu Zhiwei,Zhang Lingling
The development of a bio-sensing strategy based on CRISPR/Cas that is exceptionally sensitive is crucial for the identification of trace molecules. Colorimetric miRNA detection utilizing CRISPR/Cas13a-triggered DNAzyme signal amplification was described in this article. The developed strategy was implemented for miRNA-21 detection as a proof of concept. The cleavage activity of Cas13a was triggered when the target molecule bonded to the Cas13a-crRNA complex and cleaved uracil ribonucleotides (rU) in the substrate probe. As a consequence, the S chain was liberated from the T chain that had been modified on magnetic beads (MB). The G-rich sections were then exposed when the catalytic hairpin assembly between the H1 and H2 probes was activated by the released T@MB. G-rich section can fold into G-quadruplex. By catalyzing the formation of green ABTS 3 – via HRP-mimicking G-quadruplex/hemin complexes, colorimetric measurements of miRNA can be achieved visually through DNAzyme-mediated signal amplification. The method demonstrated a low limit of detection of 27 fM and a high selectivity towards target miRNA eventually. As a result, the developed strategy provides a clinical application platform for the detection of miRNAs that is both ultrasensitive and extremely specific.
研究领域:
生物技术
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分子生物学
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诊断方法
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DOI
PubMed
高影响因子
Aquaculture Reports
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2023
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IF 3.7
Hui Li,Shuangfeng Li,Liming Zhao,Shuying Zhang,Jing Zhang,Xianfeng Wang,Junping Liang
Cottonseed meal has been one of the predominant protein feedstuffs used in aqua feed, which contains gossypol . It is well known that gossypol, a natural polyphenolic compound found in seeds, can be toxic to fish. The toxicity of gossypol persists even after a recovery period; however, the underlying mechanism remains largely unknown. To investigate the toxic mechanism of gossypol after the recovery period, Cyprinus carpio were administered a continuous oral dose of 20 mg/kg·d gossypol (gossypol acetate dose/fish weight·d) for one week, followed by a two-week recovery period. The spleen index, spleen morphology, lysozyme (LZM) activity, acid phosphatase (ACP) activity and nitric oxide (NO) concentration were obviously changed in the spleens of C. carpio after two weeks of recovery. Then, miRNA sequencing identified 22 differentially expressed miRNAs in the spleens of C. carpio two weeks after the gossypol treatment. The mRNA expression levels of immune-associated genes were significantly increased in the spleens after recovery. MiR-194 was chosen for future study through a search of existing literature and an analysis of miRNA and mRNA expression levels. To further characterize the potential target of miR-194, we identified putative miR-194 binding sites in the 3′UTR of myeloid differentiation factor 88a (MyD88a). Dual-luciferase reporter assays revealed that miR-194 downregulated the expression level of MyD88a through binding to seed-matching sequence in the 3′ UTR of MyD88a. Moreover, C. carpio were intraperitoneally administered with miR-194 antagomir. Furthermore, down-regulation of miR-194 increased the expression levels of MyD88a and some pro-inflammatory cytokines and anti-inflammatory cytokines. Altogether, the results demonstrate miR-194 can target MyD88a to regulate the immune response of C. carpio after a two-week recovery period following gossypol administration.
研究领域:
水产养殖
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毒理学
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免疫学
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分子生物学
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产品关联
DOI
高影响因子
Scientific Reports
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2016
·Vol. 6
·期 1
·1-9
·
IF 5.228
Miao Xiangmin,Ning Xue,Li Zongbing,Cheng Zhiyuan
Positively charged gold nanoparticles (+)AuNPs can adsorb onto the negatively charged surface of single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA). Herein, long-range dsDNA polymers could form based on the hybridization chain reaction (HCR) of two hairpin probes (H 1 and H 2 ) by using miRNA-21 as an initiator. (+)AuNPs could adsorb onto the negatively charged surface of such long-range dsDNA polymers based on the electrostatic adsorption, which directly resulted in the precipitation of (+)AuNPs and the decrease of (+)AuNPs absorption spectra. Under optimal conditions, miRNA-21 detection could be realized in the range of 20 pM-10 nM with a detection limit of 6.8 pM. In addition, (+)AuNPs used here are much more stable than commonly used negatively charged gold nanoparticles ((−)AuNPs) in mixed solution that contained salt, protein or other metal ions. Importantly, the assay could realize the detection of miRNA in human serum samples.
研究领域:
纳米技术
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分析化学
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生物传感器
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分子诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2025
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IF 6
Jiabin Yang,Min Wang
Background Abnormal expression of microRNA (miRNA) is closely associated with the occurrence and development of cancer and various other diseases. Therefore, obtaining quantitative information on miRNA expression, especially detecting miRNA expression levels in living cells, is of great significance for studying related physiological and pathological processes as well as diagnosing diseases. RNA-cleaving DNAzyme (RCD)-based DNA walkers have shown great potential in the detection of miRNA. However, their application is hindered by low intracellular delivery efficiency, limited catalytic efficiency, and the requirement for additional supply of cofactors. Results In this study, we developed a novel self-feedback dual-walker (SFDW) nanosystem, loaded onto urchin-like nano-MnO 2 (UNM), for intracellular trace-level miR-21 imaging. The SFDW nanosystem incorporates a hairpin-structured capture probe and two mutually activating DNA walkers. During the detection process, the capture probe recognizes and binds to miR-21 in the matrix, opens the hairpin structure, and subsequently activates the DNA walkers. The cleavage product of each DNA walker in the nanosystem serves as the activator for the other, thereby establishing a biocircuit (self-feedback circuit) with cyclic amplification effect between the two. This mechanism significantly enhances the signal amplification capability. The UNM carrier not only promotes cellular uptake for the SFDW nanosystem, but supplies endogenous Mn 2+ cofactors via intracellular glutathione-triggered degradation as well. The SFDW nanosystem achieved a low detection limit of 2.8 pM, and successfully enabled miRNA visualization in living cells. Significance The self-feedback circuit improved detection sensitivity and reduced the detection limit, while the introduction of UNM enhanced delivery efficiency and provided the necessary cofactors. This proposed strategy can address the limitations of RCD-based DNA walkers in intracellular miRNA detection and holds great potential for clinical diagnosis and early-stage disease screening.
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2019
·
IF 9.518
Huili Shao,Han Lin,Zhiyong Guo,Jing Lu,Yaru Jia,Meng Ye,Fengmei Su,Lingmei Niu,Weijun Kang,Sui Wang,Yufang Hu,Youju Huang
MicroRNAs are widely used as tumor markers for cancer diagnosis and prognosis. Herein, a multiple signal amplification sandwich-type SERS biosensor for femtomolar detection of miRNA is reported. The signal unit consisted of giant Au vesicles, DNA sequences and deposited silver nanoparticles . The giant Au vesicles provided large-volume hot spots because of sharp tips and abundant hotspot gaps, thus enhancing the electromagnetic intensity for the SERS performance. Further silver stain would easily lead to second-stage amplification of Raman signal. In addition, more SERS signal molecules R6G adsorbed on the signal unit with the aid of HCR and the controlled nanogaps between adjacent AgNPs, brought about the third-stage amplification. The capture unit, prepared by immobilizing the capture probe (CP) on the Fe 3 O 4 @AuNPs, could easily capture target miRNA and greatly simplify the separation step to improve reproducibility. The higher concentration of target miRNA definitely formed more sandwich-type structures with combination of capture unit and signal unit, resulting in multiple amplification of SERS signals. The proposed multiple signal amplification sandwich-type SERS biosensor could detect miRNA-141 at the femtomolar level with a low detection limit of 0.03 fM. Meanwhile, it exhibited high selectivity and accuracy, even for practical analysis in human serum. Therefore, the designed multiple signal amplification sandwich-type SERS biosensor would be a very promising alternative tool for the detection of miRNA and analogs in the field of biomedical diagnosis.
研究领域:
生物医学诊断
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分子生物学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2025
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IF 10.5
Jing Peng,Simin Liu,Ziyue Qiu,Luyin Zhang,Ziyan Dong,Qianying Zhou,Ting Huang,Yanfei Zhang,Jingyu Shi,Jin-Xiang Chen,Zong Dai,Jun Chen
miRNA is an effective disease diagnostic biomarker. Developing efficient miRNA in living cells analysis methods, holds significant importance for disease diagnosis and understanding disease progression. Strategies combining DNA nanostructures and enzyme-free amplification have been developed for highly sensitive and rapid imaging of miRNA. However, due to nonspecific amplification of enzyme-free amplification inherent in the probe assembly process and during transfection, these strategies suffer from significant background signals and reduced sensitivity, limiting their further application. To overcome these limitations, we have developed a DNA tetrahedral-based and photocontrolled intramolecular CHA strategy (TCHA-P) to achieve spatiotemporal, highly sensitive, and accurate imaging of miRNA in living cells. In this strategy, the DNA tetrahedron not only serves as a transfection vector but also enhances the local concentration of reactant probes, boosting the reaction kinetics and sensitivity of CHA. Additionally, the light-controlled group (PC-linker) can trigger CHA at selected times and locations, enhancing the spatiotemporal precision of miRNA imaging and ensuring high sensitivity and accuracy by eliminating nonspecific amplification during the assembly and transfection processes. Using miR-10b as a proof-of-concept model, the proposed strategy demonstrated higher signal-to-noise ratio compared to traditional CHA, with a detection limit of 28 pM. Living cell imaging experiments showed that the strategy could effectively differentiate the expression of miR-10b between normal breast cells and breast cancer cells, providing high imaging contrast. The strategy holds promise for broadening the path of nucleic acid amplification-based imaging and is expected to become an effective tool in nucleic acid-based disease diagnosis, treatment, and drug development.
研究领域:
分子生物学
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纳米生物学
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医学诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Yang Lu,Yiyue Yu,Yeru Wang,Wenbo Zhou,Ziyi Cheng,Liandong Yu,Shaojiang Zheng,Rongke Gao
Background Precise and specific miRNA detection plays a vital role in exploring development mechanisms of cancer disease, thereby it can significantly improve relevant prevention and treatment strategies. Results In this work, a surface-enhanced Raman spectroscopy (SERS)-based microfluidic chip has been devised with a microcone array SERS substrate (MCASS) for the miR-141 detection. This substrate excels in unique SERS activity and large surface area for DNA oligonucleotide modification. As the presence of miR-141, the DNAzyme walker induced cleavage reaction took place on the finely designed and prepared dual DNA conjugated SERS nanoprobes. The SERS nanoprobes can anchor on MCASS by the DNA hybridization that achieved an impressive detection limit in the femtomolar level. Significance With this integrated SERS-based microfluidic chip, we provided a miRNA detection strategy using DNAzyme walker amplification technology. It is believed that this strategy could be a powerful tool for miRNA detection and related cancer screening test .
研究领域:
生物医学工程
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分子生物学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
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2023
·Vol. 190
·期 4
·1-9
·
IF 5.7
Su Zhanying,Zhang Li,Yu Ying,Lin Bixia,Wang Yumin,Guo Manli,Cao Yujuan
An electrochemical strategy based on bimetallic nanozyme in collaboration with toehold-mediated DNA replacement effect is proposed for the sensitive determination of miRNA-21. The AuPt nanoparticles (AuPt NPs) are prepared as a catalytic beacon; it shows favorable peroxidase properties with a Michaelis contant ( K m ) of 0.072 mM for H 2 O 2 , which is capable of catalyzing H 2 O 2 to induce an intense redox reaction, and causing a measurable electrochemical signal. To further enhance the strength of the signal response, a novel toehold-mediated DNA replacement strategy is employed. DNA strands with specific sequences are modified on electrodes and AuPt NPs, respectively. In the presence of miRNA-21, a cyclic substitution effect is subsequently activated via a specific toehold sequence and leads to a large accumulation of AuPt NPs on the electrodes. Subsequently, a strong signal depending on the amount of miRNA-21 is obtained after adding a small amount of H 2 O 2 . The analytical range of this determination method is from 0.1 pM to 1.0 nM, and the LOD is 84.1 fM. The spike recoveries for serum samples are 95.0 to 102.4% and the RSD values are 3.7 to 5.8%. The results suggests a promising application of the established method in clinical testing and disease diagnosis. Graphical Abstract
研究领域:
生物化学
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分子诊断
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纳米医学
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产品关联
DOI
PubMed
高影响因子
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
·
2024
·
IF 3.8
Ying Zhang,Jing Xie,Huimin Zhang,Jiacheng Li,Xing Mi,Xuyi Zhou,Zhenhua Ding
Introduction Local exposure to ionizing radiation (IR) can induce changes in biological processes in distant tissues and organs. Exosomes are nanoscale vesicles that transport biomolecules, mediate communication between cells and tissues, and can affect the abscopal effects of radiotherapy. Methods Mice were treated with 8.0 Gy doses of chest and abdomen IR, after which serum samples were taken 24 h after exposure. Their serum exosomes were then isolated via ultracentrifugation and the small RNA portions were extracted for sequencing and bioinformatic analysis. Exosomes were injected intravenously into the mice to assess their ability to cross the blood-brain barrier (BBB). Glioma cells and gliomal stem cells (GSCs) were examined for malignant biological behaviors, stemness, and tumorigenic capacity after co-culturing with different types of exosomes. Results We found that serum exosomes crossed the BBB in mice after local IR exposure—which induced decreases in the expression of BBB tight-junction proteins and increased brain endothelial cell apoptosis. Exosomes from the exposed groups promoted malignant biological behaviors, stemness, and tumorigenic capacity in glioma cells and GSCs by upregulating the expression of SOS1. Phospho-MEK1/2 and Phospho-ERK1/2, of the MAPK signaling pathway, were found to be up-regulated in cells that were co-cultured with the exposing groups of the exosomes. Further analyses demonstrated that differentially expressed levels of miR-93-5p in mouse serum exosomes regulated the cellular expression of SOS1. Conclusion Following local IR exposure, serum exosomes cross the BBB to promote the progression of distant gliomas. Exosomal microRNAs play an important role in this process.
研究领域:
肿瘤学
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分子生物学
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放射生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2022
·
IF 8.008
Yudie Sun,La Fang,Yunxiang Han,Aobo Feng,Shengjun Liu,Kui Zhang,Jing-Juan Xu
Myocardial miRNAs in peripheral blood are closely related to the pathogenic process of myocardial infarction. Rapid identification and accurate quantification of myocardial miRNAs are of great significance to clinical interventions for treating cardiovascular lesions. Therefore, a ratiometric electrochemiluminescence (ECL) biosensor integrating DNAzyme with a resonance energy transfer (RET) system was designed to detect myocardial miRNA. The dual-signal system was composed of rA marked substrate strand functionalized CdTe quantum dots (QDs) as reductive–oxidative (R–O) emitters and Cy5-labeled strand-functionalized Ru(bpy)32+-filled silica nanoparticles (RuSi NPs) as oxidative–reductive (O–R) emitters. In the presence of target miRNA, DNAzyme was activated to cut substrate strands on the CdTe QDs and release triggers for opening hairpin probes. Then, the Cy5 molecule-labeled hairpin DNA on the RuSi NPs was opened to introduce Cy5 molecules and RuSi NPs into the system. The R–O ECL was quenched by ECL-RET between CdTe QDs and Cy5 molecules and the O–R ECL was introduced by the RuSi NPs. In this way, based on the simultaneous changing of the ECL signal, the dual-potential dynamic signal ratiometric ECL sensing platform was developed. By measuring the ratio of O–R ECL signal to R–O ECL signal, the concentration of miRNA-499 was accurately quantified in the range of 10 fM to 10 nM, and the detection limit was as low as 2.44 fM (S/N = 3). This DNAzyme guided dual-potential ratiometric ECL method provides a sensitive and reliable method for myocardial miRNA detection, and it has great potential in clinical diagnosis and treatment.
研究领域:
分析化学
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生物传感器
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心脏病学
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纳米技术
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2022
·Vol. 14
·期 44
·4523-4530
·
IF 3.532
Jiao Wang,Liyuan Wen,Ruyuan Cao,Xiaorong Gao,Xia Li,Ensheng Xu,Qi Zhang,Shuling Xu,Caifeng Dai,Qingwang Xue
Single miRNAs are not specific and accurate enough to meet the strict diagnosis requirements in practice. Therefore, simultaneous monitoring of multiplexed miRNA in biological samples can not only improve the accuracy and specificity of bioassays but also avoid the squandering of valuable biological specimens. Herein, we designed a metal sulfide nanoparticle-based dual barcode-triggered DNAzyme cascade strategy for the sensitive and simultaneous multiplex miRNA detection in a single assay. Firstly, the capture probes (H1, H2) specifically recognize targets (miRNA-21, miRNA-141), exposing the stem of H1 and H2. Then, with the introduction of a detection probe (CuS-H3, ZnS-H4), the exposed H1 and H2 catalyze the hairpin assembly (CHA) reaction, realizing target miRNA recycling, and forming H1/H3-CuS and H2/H4-ZnS complexes. Subsequently, the formed H1/H3-CuS and H2/H4-ZnS complexes are encoded on magnetic beads through the biotin/streptavidin interaction. The CuS and ZnS nanoparticles captured by magnetic beads release thousands of Cu2+ and Zn2+via the cation exchange reaction. Finally, the released Cu2+ and Zn2+ specially activate the DNAzyme of the catalytic and molecular beacon (CAMB) system. The CAMB system affords an amplified fluorescence signal output by cycling and regenerating the metal ion-dependent DNAzyme to realize multiple enzymatic turnovers. Benefiting from target recycling, nanoparticle amplification, and catalytic and molecular beacon amplification, there is substantial amplification and the target miRNAs can be detected at 0.06 fM (miRNA-21) and 0.048 fM (miRNA-141) in a single assay. Furthermore, the high selectivity and accuracy of the assay were proved by practical analysis of different cancer cells, which exhibited good practicability in multiplex miRNA detection in clinical sera. The results indicate that the proposed strategy holds great potential for the sensitive detection of multiplex cancer biomarkers and offers the opportunity for future applications in clinical diagnosis.
研究领域:
生物化学
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分子诊断
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纳米医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Sensors and Actuators Reports
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2025
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IF 7.6
Haoyue Ma,Ying Liang,Jiangnan Wang,Wanjuan Lin,Yang Pan,Peng Cao,Yue Zhang
The development of colorimetric biosensors offers a promising tool for on-site visual analysis of medicinal plants and related products in market regulation. However, compared to fluorescent or electrochemical sensing strategies, colorimetric sensors often suffer from weak signals and limited sensitivity, making it challenging to detect low-abundance RNA targets in commercial samples. In response to this issue, here we design a triple signal amplification strategy to achieve ultrasensitive colorimetric signal output for visible microRNA (miRNA) detection in medicinal plants and products, and the smartphone-based imaging enabled portable quantitative analysis for on-site market regulation. The specifically expressed miRNA in ginseng is chosen as the target strand to initiate the first amplification (hybridization chain reaction) for DNAzyme activation, catalyzing the successive cleavage of substrate strands for the liberation of trigger strands that drive rolling circle amplification. This process produces a high abundance of repetitive G-quadruplex motifs, which function as catalytic scaffolds to accelerate a colorimetric reaction, ultimately enabling sensitive and reliable detection by the naked eye. Combining three amplification approaches into a unified cascade, this strategy generates distinguishable colorimetric signals with naked eyes, and the quantitative analysis could be achieved using a smartphone software, offering a wide dynamic range (1-10 5 pM) and a low detection limit of 651 fM. Successful application in medicinal plant ginseng and related products demonstrates the practicality for real sample analysis. The modular amplification design holds broad potential for detecting other molecular targets, underscoring its promise for portable biosensing and on-site regulatory applications.
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DOI
高影响因子
危险品化学品经营许可证（带存储）