Chemical Science
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2017
·Vol. 9
·期 5
·1184-1190
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IF 8.668
Xiangdan Meng,Wenhao Dai,Kai Zhang,Haifeng Dong,Xueji Zhang
Herein, we design a smart autonomous ATP self-powered strand-displacement cascade amplification (SDCA) system for highly sensitive multiple intracellular miRNA detection. Rationally engineered Y-motif DNA structures are functionalized on mesoporous silica-coated copper sulfide nanoparticles loaded with numerous ATPs (CuS@mSiO2-Y/ATP) through pH stimulus-responsive disulfide bonds. The SDCA system is implemented by endogenous specific miRNA as a trigger and ATP as fuel released from the nanocarrier at acidic pH and photothermal stimuli-responsive CuS. The ATP self-powered SDCA process presents higher sensitivity compared to that without amplification for intracellular miRNA imaging. Two-color simultaneous and sensitive imaging of multiple cancer-related miRNAs in living cells is also confirmed. This enables facile and accurate differentiation between normal cells and different types of cancer cell using intracellular miRNA imaging, which improves the veracity and timeliness for early cancer diagnosis.
研究领域:
纳米技术
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生物医学工程
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分子生物学
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癌症研究
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2019
·
IF 6.35
Yun Ying Cheng,Yi Fen Xie,Chun Mei Li,Yuan Fang Li,Cheng Zhi Huang
Förster resonance energy transfer (FRET) by using fluorescent carbon dots (CDs) as energy donors shows potential for biosensing and bioimaging. However, it remains underused and underestimated for CDs as a building block for FRET owing to the low efficiency and complex operation originating from the surface modification of CDs. To overcome these limitations, herein we develop a novel FRET soft nanoball (fretSNB) in which thousands of green CDs and black hole quencher 2 (BHQ-2) dyes are loaded, and FRET occurs from CDs to BHQ-2 dyes with the consequence of effective fluorescence quenching. These fretSNBs can be ruptured in the presence of phospholipase A2 (PLA2) released in a process of duplex-specific nuclease (DSN)-assisted target recycling amplification (TRA), making the fluorescence of CDs recovered. Thus, a dual amplification strategy is successfully developed for amplified detection of microribonucleic acids (miRNAs) in the concentration range 0.025–10 nM with a limit of detection (3σ) reaching 16.5 pM which is about 515 times lower than without fretSNBs. In addition, the developed strategy exhibits high selectivity for discrimination of a single nucleotide difference and capability to detect miRNAs extracted from cells, suggesting excellent potential in biomedical analysis and clinical diagnosis.
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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生物医学工程
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DOI
PubMed
高影响因子
ACS Sensors
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2021
·
IF 7.711
Wenjun Zhong,Qiufang Yang,Kerui Fang,Dan Xiao,Cuisong Zhou
The simultaneous discrimination of let-7 microRNAs (miRNAs) would greatly facilitate the early diagnosis and prognosis monitoring of diseases. In this work, a molecular beacon DNA probe was designed to be able to flip out its mismatched cytosine base within the α-hemolysin (α-HL) latch and generate completely separated blocking currents to identify the single-base difference. As a result, the characteristic blocking current of fully matched MB/let-7a and single-base mismatched MB/let-7f was 84.30 ± 0.92 and 87.05 ± 0.86% (confidence level P 95%), respectively. Let-7 miRNA family let-7a and let-7f were completely simultaneously discriminated, which could be attributed to the following strengths. (1) The statistic distribution of blocking current is extremely concentrated with a small relative standard deviation (RSD) of less than 1% and a narrow distribution range. (2) Complete separation is achieved with a high separation resolution of 1.54. (3) The cytosine base flipping out within the α-HL latch provides a universal labeling-free strategy to simultaneously discriminate the single-base mismatch. Overall, the target let-7f sequences were detected with a linear range from 0.001 to 10 pM in human serum samples containing 200 nM let-7a. Great potential has been demonstrated for precise detection, early diagnosis, and prognosis monitoring of diseases related to single-base difference.
研究领域:
纳米技术
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生物物理学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
Journal of the American Heart Association
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2022
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IF 6.106
Xiaofei Luo,Chengcheng Hang,Ziming Zhang,Kaixing Le,Yuhan Ying,Ying Lv,Lingling Yan,Yajie Huang,Lixia Ye,Xuefeng Xu,Ying Zhong,Lizhong Du
BackgroundIntrauterine growth restriction (IUGR) is closely related to systemic or pulmonary hypertension (PH) in adulthood. Aberrant crosstalk between pulmonary vascular endothelial cells (PVECs) and pulmonary arterial smooth muscle cells (PASMCs) that is mediated by exosomes plays an essential role in the progression of PH. FoxM1 (Forkhead box M1) is a key transcription factor that governs many important biological processes.Methods and ResultsIUGR‐induced PH rat models were established. Transwell plates were used to coculture PVECs and PASMCs. Exosomes were isolated from PVEC‐derived medium, and a microRNA (miRNA) screening was proceeded to identify effects of IUGR on small RNAs enclosed within exosomes. Dual‐Luciferase assay was performed to validate the predicted binding sites of miRNAs on FoxM1 3’ untranslated region. FoxM1 inhibitor thiostrepton was used in IUGR‐induced PH rats. In this study, we found that FoxM1 expression was remarkably increased in IUGR‐induced PH, and PASMCs were regulated by PVECs through FoxM1 signaling in a non‐contact way. An miRNA screening showed that miR‐214‐3p, miR‐326‐3p, and miR‐125b‐2‐3p were downregulated in PVEC‐derived exosomes of the IUGR group, which were associated with overexpression of FoxM1 and more significant proliferation and migration of PASMCs. Dual‐Luciferase assay demonstrated that the 3 miRNAs directly targeted FoxM1 3’ untranslated region. FoxM1 inhibition blocked the PVECs‐PASMCs crosstalk and reversed the abnormal functions of PASMCs. In vivo, treatment with thiostrepton significantly reduced the severity of PH.ConclusionsTransmission of exosomal miRNAs from PVECs regulated the functions of PASMCs via FoxM1 signaling, and FoxM1 may serve as a potential therapeutic target in IUGR‐induced PH.
研究领域:
心血管疾病
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分子生物学
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细胞生物学
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转化医学
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DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 8
·1-9
·
IF 5.3
Zhou Yanmei,Che Shengjun,Wang Zhili,Zhang Xiaoru,Yuan Xunyi
An electrochemical sensor assisted by primer exchange reaction (PER) and CRISPR/Cas9 system (PER-CRISPR/Cas9-E) was established for the sensitive detection of dual microRNAs (miRNAs). Two PER hairpin (HP) were designed to produce a lot of extended PER products, which could hybridize with two kinds of hairpin probes modified on the electrode and initiate the cleavage of two CRISPR/Cas9 systems guided by single guide RNAs (sgRNAs) with different recognition sequences. The decrease of the two electrochemical redox signals indicated the presence of dual-target miRNAs. With the robustness and high specificity of PER amplification and CRISPR/Cas9 cleavage system, simultaneous detection of two targets was achieved and the detection limits for miRNA-21 and miRNA-155 were 0.43 fM and 0.12 fM, respectively. The developed biosensor has the advantages of low cost, easy operation, and in-situ detection, providing a promising platform for point-of-care detection of multiple miRNAs. Graphical abstract
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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生物技术
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Fang Hu,Wancun Zhang,Jiaqi Zhang,Qi Zhang,Tao Sheng,Yueqing Gu
In this study, a simple, economic, and label-free electrochemical biosensor was developed for highly sensitive and selective miR-21 detection based on Fe(CN) 6 3− /Fe(CN) 6 4− electrochemical signal, which relies on DNA structures conjugating with carboxyl multi-wall carbon nanotubes (MWCNTs-COOH)-modified glassy carbon electrode (GCE). High sensitivity and specificity of the biosensor were achieved by taking advantage of the target-recycled non-enzymatic amplification strategy (TRNEAS), which relies on sequence-specific hairpin strand displacement and does not require the addition of environment-susceptible enzymes. Besides, the SP-DNA strictly binds to the cMWCNTs/GCE via amide bonds, which ensure good electrochemical results and excellent stability for target miRNA detection. The developed miR-21 electrochemical biosensor exhibited a broad linear dynamic range of 0.1 fmol to 5 pmol and a detection limit of 56.7 amol for miR-21 detection in vitro . The relative miR-21 expression levels were particularly detected by our developed miR-21 biosensor and were highly consistent with the stem-loop RT-PCR results. In summary, our developed electrochemical biosensor exhibits great potential for further application in biomedical research and early clinical diagnosis.
研究领域:
生物传感器
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电化学
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纳米技术
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分子生物学
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临床诊断
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DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2022
·
IF 9.221
Fu-Ting Wang,Hu Luo,Yang-Yang Hou,Yu Ya,Ke-Jing Huang,Shu-Peng Li,Lan-Rui Wang,Ya-Lin Song,Xuecai Tan
We present a dual-microRNAs-driven enzyme biofuel cells (EBFCs) based visual self-powered biosensing system with a capacitor assistance significant amplification of the output signal for the sensitive detection of microRNA-21 and microRNA-141. When microRNA-21 is captured by the aptamer (Atp1–21), the glucose oxidase (GOD)-modified aptamer (Atp2–21-GOD) is immobilized on the electrode surface by base-complementary pairing. When microRNA-21 and microRNA-141 are captured by the dual aptamer recognition system, the enzyme load is more effectively increased and the input voltage of the EBFCs is significantly enhanced. In the linear range of 0.0005–100 PM and 0.001–1000 PM, the detection limits for microRNA-21 and microRNA-141 are 0.16 and 0.36 fM, respectively. Furthermore, the EBFCs are connected to the capacitor integrated circuit, and the input voltage of the EBFCs is converted to an enhanced instantaneous current and charging to the capacitor which is then directly detected by a multimeter , and the sensitivity of sensing system is improved by 29.48 times. In order to improve the reproducibility and accuracy of self-powered biosensing system, the reading of the multimeter is obtained in real time by software that opens new door to the visualization of detection signal. The method provides an efficient strategy for visualization of multimeter-based detection signal and opens the possibility for medical diagnosis and therapeutic device power in the biosensing device.
研究领域:
生物化学
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分子生物学
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医学诊断
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Pei Wang,Xiaowei Wei,Luming Shen,Kexin Xu,Zhongting Wen,Nengjiao Gao,Ting Fan,Shenmei Xun,Qingyi Zhu,Xiaojun Qu,Yefei Zhu
Bladder cancer (BC) occurrence and progression are accompanied by alterations in microRNAs (miRNAs) expression levels. Simultaneous detection of multiple miRNAs contributes to the accuracy and reliability of the BC diagnosis. In this work, wrinkled silica nanoparticles (WSNs) were applied as the microreactor for multiplex miRNAs analysis without enzymes or nucleic acid amplification. Conjugated on the surface of WSNs, the S9.6 antibody was adopted as the universal module for binding DNA/miRNA duplexes, regardless of their sequence. Furthermore, single-stranded DNA (ssDNA) was labeled with quantum dots (QDs) for identifying a given miRNA to form QDs-ssDNA/miRNA, which enabled the specific capture of the corresponding QDs on the wrinkled surface of WSNs. Based on the detection of fluorescence signals that were ultimately focused on WSNs, target miRNAs could be sensitively identified to a femtomolar level (5 fM) with a wide dynamic range of up to 6 orders of magnitude. The proposed strategy achieved high specificity to obviously distinguish single-base mutation sequences and possessed multiplex assay capability. Moreover, the assay exhibited excellent practicability in the multiplex detection of miRNAs in clinical serum specimens.
研究领域:
癌症生物学
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纳米技术
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分子诊断
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2024
·
IF 4.3
Lin Liu,Jia Hui He,Xiao Qiao Wu,Jia Jun Liu,Wen Yi Lv,Cheng Zhi Huang,Hui Liu,Chun Mei Li
MicroRNAs (miRNAs) play a key role in physiological processes, and their dysregulation is closely related to various human diseases. Simultaneous detection of multiple miRNAs is pivotal to cancer diagnosis at an early stage. However, most multicomponent analyses generally involve multiple excitation wavelengths, which are complicated and often challenging to simultaneously acquire multiple detection signals. In this study, a convenient and sensitive sensor was developed to simultaneously detection of multiple miRNAs under a single excitation wavelength through the fluorescence resonance energy transfer between the carbon dots (CDs)/quantum dots (QDs) and graphene oxide (GO). A hybridization chain reaction (HCR) was triggered by miRNA-141 and miRNA-21, resulting in the high sensitivity with a limit of detection (LOD) of 50 pM (3 σ/k ) for miRNA-141 and 60 pM (3 σ/k ) for miRNA-21. This simultaneous assay also showed excellent specificity discrimination against the mismatch. Furthermore, our proposed method successfully detected miRNA-21 and miRNA-141 in human serum samples at a same time, indicating its diagnostic potential in a clinical setting.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
PubMed
高影响因子
iScience
·
2023
·
IF 5.8
Huo Xu,Fenglin Yang,Danlong Chen,Weilin Ye,Guohui Xue,Lee Jia
MicroRNAs (miRNAs) play an important biomarker in various biological processes, especially cancer related, yet economic, simple, sensitive and specific methods for miRNA determination are still challenging. In this study, we have developed stepwise-strand displacement amplification (S-SDA)-based colorimetric sensing platform for let-7a miRNA detection in clinical serum samples. Our results demonstrated that the developed S-SDA-based method shows high sensitivity with a detection limit of 63.2 pM and a naked eye detection limit of 0.1 nM. Moreover, the S-SDA amplifier is able to discriminate target miRNAs from their mutants with high accuracy and specificity. With its high sensitivity and selectivity, this method successfully identified healthy individuals from patients with colon cancer by detecting let-7a miRNAs in serum. We believe the colorimetric analysis method will provide a new paradigm for the detection of miRNA with different abundance and show great potential for clinical application in biomedical analysis and early clinical diagnosis.
研究领域:
生物化学
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分子生物学
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临床诊断
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DOI
PubMed
高影响因子
Nanoscale
·
2019
·Vol. 11
·期 36
·16879-16885
·
IF 6.97
Han Ding,Yi An,Tao Zhao,Bing Liu,Yin Wang,Lei Zhang,Yu Wang,Yuan Zhang,Man Wang,Yanhan Dong,Longgang Hu,Bu-Chang Zhao,Peifeng Li
microRNAs are a type of evolutionarily conserved small non-coding RNA with a length of 18–25 nucleotides. In recent years, increasing studies have shown that the content of specific miRNAs in the blood changes significantly during the occurrence and development of major diseases such as cardiovascular disease and cancer. Therefore, miRNAs may serve as important new biomarkers that can be used for disease diagnosis in the future. Here, we improved the polyethylene glycol layer on the surface of a traditional silicon sphere to specifically capture miRNAs by means of a full-function microplate detector, at 100 microliters. The detection limit for specific miRNAs per liter of plasma can reach 1 fM, and simultaneous detection of 96 samples can be achieved. Compared with the traditional real-time PCR technology, our detection eliminates the complex steps of miRNA extraction, reverse transcription, amplification, etc. and avoids more human error in the detection process. Using the full-featured microwell detector, we can rapidly detect specific miRNAs in plasma, which can be used in the diagnosis of cardiovascular diseases in the future.
研究领域:
生物标志物研究
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医学诊断
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分子生物学
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生物技术
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2020
·
IF 3.807
Xialing Hou,Zhiguang Suo,Ziheng Hu,Xinying Zhang,Yingying Chen,Lingyan Feng
A label-free microRNAs (miRNAs) electrochemiluminescence sensor was developed based on the synergistically enhanced ECL signal using novel ECL nanomaterial assembled by tri-luminophores perylenetetracarboxylic acid (PTCA) graphite-like carbon nitride nanosheet (g-C 3 N 4 NS) and graphene carbon quantum dots (GQDs). The ECL intensity of our proposed nanocomposites increased significantly, by taking advantages of the synergistic promotion ECL mechanism of PTCA g-C 3 N 4 and GQDs. Meanwhile, a three-way junction (TWJ) structure was introduced into the sensor design, whose conformation can be specifically induced in the presence of target miRNA. Thus, the formation of TWJ structure can hinder the contact between the co-reactant Na 2 S 2 O 8 in the solution phase and the luminescent substrate, which produce a quenched ECL signal. Our proposed ECL biosensor provided a highly sensitive detection of miRNA-21 with a detection limit of 96 aM ranging from 100 aM to 1 nM. Furthermore, this ECL sensing system was able to successfully distinguish the target RNA from its homologous family and presented a promising response in human real samples. With excellent properties, the proposed ECL biosensor provided an efficient and ultrasensitive method for miRNAs detection.
研究领域:
分析化学
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生物化学
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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
高影响因子
危险品化学品经营许可证（带存储）