JOURNAL OF MEDICINAL CHEMISTRY
·
2023
·
IF 7.3
Zhen Song,Qiang-Yan Zhang,Jia-Jing Li,Jing-Lian Su,Yuan-Hua Liu,Gong-Jun Yang,Huai-Song Wang
Let-7a, a type of low-expressed microRNAs in cancer cells, has been investigated as a promising biomarker and therapeutic target for tumor suppression. Developing simple and sensitive detection methods for let-7a is important for cancer diagnosis and treatment. In this work, the hybridization chain reaction (HCR) was initiated by let-7a via two hairpin primers (H1 and H2). After the HCR, the remaining hairpin H1 was further detected by lateral flow assay (LFA) and electrochemical impedance spectroscopy. For LFA, biotin-modified H1(bio-H1) and free H2 were used for HCR. With the decrease of let-7a concentration, the color of T line gradually increased. As for electrochemical methods, the H1′-AuNP-modified electrode was used for detection of bio-H1 based on the difference of impedance (ΔRct) detected without and with different concentrations of let-7a participating in the HCR. This method could detect let-7a in the range of 10.0 fM and 1.0 nM with detection limits of 4.2 fM.
研究领域:
癌症诊断
·
分子生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Kai Zhang,Zhenqiang Fan,Yue Huang,Minhao Xie,Jianfeng Zhao,Jiaying Wang
We have carefully designed a probe for in situ detection and imaging of Argonaute2 (Ago2), a key RNA interference (RNAi) protein in human cells and siRNA (Let-7a) interactions. The probe mainly contains two parts: a gold nanoparticle (AuNP) being quenched and a functionalized nucleic acid as a fluorescent output carrier. After the probe is endocytosed into the cell, the modified hairpin nucleic acid can be cut into two parts by the Ago2/Let-7a complex (ALC). The cut nucleic acid will replace a DNA strand modified on another duplex nucleic acid. The replaced DNA strand’s 3’-end is modified with a fluorophore (Cy3). By cascading single cells, the probe can directly achieve the fluorescence recovery of the probe in the cell. By dynamically monitoring the in-situ quantitative response of the interaction between Ago2 and Let-7a, and distinguishing between strongly RNA-expressing cells and other cells, the interaction between Ago2 and Let-7a can be monitored dynamically, proving the utility of the probe. Probes will accelerate the basic role of reducing protein-RNA interactions, the mystery of RNA silencing, and monitoring the treatment of cancer.
研究领域:
分子生物学
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细胞生物学
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生物技术
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癌症研究
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产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2022
·
IF 6.911
Wenwen Xue,Zilian Jiang,Yahui Wang,Hongfang Zhang
Sensitive and accurate detection of tumor suppressor genes is vastly important to the related therapeutic research. Herein, a ratiometric electrochemical method for let-7a detection was established by integrating a ferrocene (Fc) doped MoS 2 nanoplates modified electrode into the nanochannels-based biosensing platform. The ratiometric signal was developed by the redox current of methylene blue (MB) which reflects the target recognition occurred into the nanochannels and the redox current of Fc which corrects the slight signal deviation caused by some analyte-independent factors. And thus, the ratio of peak current of MB and Fc ( I MB / I Fc ) measured at differential pulse voltammogram varied precisely with the increment of the concentration of let-7a incubated in the bioinspired nanochannels. The strategy of spherical DNAzyme induced deposition in nanochannels was utilized to further amplify the signal. Under optimal conditions, a wide linear dynamic range of 50 aM to 10 pM spanning five orders of magnitude was obtained. The developed electrochemical method, with attomole level of detection limit, was successfully applied to the determination of let-7a in human serum and tumor cells. The study not only offers a new route for reliable nucleic acid detection, but also provides an excellent opportunity to extend the application of the two-dimensional transition-metal dichalcogenides.
研究领域:
生物化学
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纳米技术
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电化学
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癌症研究
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分子生物学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 2
·1-8
·
IF 6.232
Li Ruiying,Liu Qiang,Jin Yan,Li Baoxin
The authors have incidentally found that the three tandem repeats of a 13-mer G-rich oligomer (with sequence 5′-TGG GAA GGG AGG G-3′; referred to as G3) can directly fold into a stable G3 trimer. The G3 trimer/hemin DNAzyme exhibits an about 3-fold higher peroxidase-mimicking activity compared to the conventional G3/hemin DNAzyme. Combining this finding with rolling circle amplification (RCA), a colorimetric assay was developed for sensitive and specific determination of microRNA. In this method, each cycle of RCA generates three catalytic units. This leads to a significant signal amplification of the RCA. Using let-7a as a model analyte, the colorimetric method (best performed at 420 nm) exhibits high sensitivity toward microRNA-let-7a with a 37 fM detection limit and an analytical range that covers 3 orders of magnitude. The method was applied to the determination of let-7a in some cell lysates.
This G-triplex trimer-based rolling circle amplification (RCA) method can produce three catalytic units per RCA cycle, which can significantly improve the amplification efficiency of RCA.
研究领域:
生物化学
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分子生物学
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纳米技术
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2012
·
IF 4.555
Sai Bi,Yangyang Cui,Li Li
Considering the great significance of microRNAs (miRNAs) in cancer detection and typing, the development of sensitive, specific, quantitative, and low-cost methods for the assay of expression levels of miRNAs is desirable. We describe a highly efficient amplification platform for ultrasensitive analysis of miRNA (taking let-7a miRNA as a model analyte) based on a dumbbell probe-mediated cascade isothermal amplification (DP-CIA) strategy. The method relies on the circularization of dumbbell probe by binding target miRNA, followed by rolling circle amplification (RCA) reaction and an autonomous DNA machine performed by nicking/polymerization/displacement cycles that continuously produces single-stranded G-quadruplex to assemble with hemin to generate a color signal. In terms of the high sensitivity (as low as 1 zmol), wide dynamic range (covering 9 orders of magnitude), good specificity (even single-base difference) and easy operation (one probe and three enzymes), the proposed label-free assay is successfully applied to direct detection of let-7a miRNA in real sample (total RNA extracted from human lung tissue), demonstrating an attractive alternative for miRNA analysis for gene expression profiling and molecular diagnostics, particularly for early cancer diagnosis.
研究领域:
分子生物学
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医学诊断
·
生物化学
✓ 已验证
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
·
临床诊断
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2016
·
IF 7.476
Hao Wu,Yaling Liu,Hongyong Wang,Jun Wu,Feifan Zhu,Pei Zou
In this study, a simple, label-free, and enzyme-free colorimetric biosensor has been developed for amplified detection of let-7a microRNA (miRNA) on the basis of dual signal amplification strategy. The sensing system mainly consists of four unlabeled hairpin probes termed H1, H2, H3, and H4. Upon sensing of the target miRNA, hairpin H1 is opened. Then hairpin H2 hybridizes with H1 forming H1–H2 duplex and frees the target miRNA that can be recycled to trigger another reaction cycle. In addition, the newly formed H1–H2 duplex hybridizes with hairpin H3, and this triggers the autonomous cross-opening of the two hairpins H3 and H4 through hybridization chain reaction. During this process, numerous split G-quadruplex structures are generated and further associate with cofactor hemin to form massive peroxidase-mimicking DNAzymes. The resulting DNAzymes catalyze the H 2 O 2 -mediated oxidation of colorless 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS 2− ) to the green-colored ABTS •− , inducing a remarkably amplified colorimetric signal. This newly developed sensing system exhibits high sensitivity toward miRNA with a detection limit of 7.4 fM and a large dynamic range of 6 orders of magnitude from 10 fM to 10 nM. Furthermore, it exhibits a good performance to discriminate single-base difference among the miRNA family members and holds a great potential for early diagnosis in gene-related diseases.
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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纳米技术
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产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2021
·
IF 9.229
Xunxun Deng,Xiaobo Liu,Shuo Wu,Shiyu Zang,Xiaotong Lin,Yanqiu Zhao,Chunying Duan
Accurate and sensitive fluorescence imaging of intracellular miRNA is essential for understanding the mechanism underlying some physiological and pathological events, as well as the prevention and diagnosis of diseases. Herein, a highly sensitive ratiometric fluorescent nanoprobe for intracellular miRNA imaging was fabricated by integrating a Ru-SiO2@polydopamine (Ru-SiO2@PDA) nanoplatform with a near-infrared light (NIR)-assisted DNA strand displacement signal amplification strategy. The Ru-SiO2@PDA spheres have excellent biosafety, high photothermal effect, and unique photophysical properties that can both emit a stable red fluorescence and well quench the fluorophores getting closer to them. So, when the fuel DNA and carboxyfluorescein (FAM)-labeled signal DNA are co-assembled on their outer surfaces, the FAM’s green fluorescence is quenched, and a low ratiometric signal is obtained. However, in the presence of miRNA, the target displaces the signal DNA from the capture DNA, releasing the signal DNA far away from the Ru-SiO2@PDA. Then, the green fluorescence recovers and leads to an enhanced Igreen/Ired value. Under NIR light irradiation, the Ru-SiO2@PDA increases the local temperature around the probe and triggers the release of fuel DNA, which thus recycles the target miRNA and effectively amplifies the ratiometric signal. Using A549 cells as a model, the nanoprobe realizes the highly sensitive ratiometric fluorescence imaging of miRNA let-7a, as well as its in vivo up- and down-regulation expressions. It provides a facile tool for highly sensitive and accurate intracellular miRNA detection through one-step incubation and may pave a new avenue for single-cell analysis.
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
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
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
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纳米技术
·
分子生物学
·
药物发现
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产品关联
DOI
PubMed
高影响因子
LUMINESCENCE
·
2020
·Vol. 35
·期 6
·835-844
·
IF 1.855
Zheng Chang,Jing Feng,Xingwang Zheng
In this paper, a convenient reverse-phase microemulsion method for the synthesis of SiO 2 nanoparticles (NPs) by simply introducing the chitosan and fluorescent dye of lucigenin during the formation reaction of SiO 2 NPs was proposed. Addition of chitosan can make the SiO 2 NPs porous, and increases lucigenin molecule incorporation into chitosan/SiO 2 NPs nanopores based on electrostatic interaction and supermolecular forces. Therefore, fluorescence quantum yield of the lucigenin/chitosan/SiO 2 composite nanoparticles was increased by introduction of chitosan and compared with lucigenin/SiO 2 NPs without chitosan. Because the number of negative charges carried when using single-stranded DNA (ssDNA) was different from that of double-stranded DNA (dsDNA), the numbers of lucigenin/chitosan/SiO 2 composite nanoparticles with positive charge adsorbed using ssDNA or dsDNA were different. Consequently, fluorescence intensity caused using ssDNA or dsDNA/miRNA was clearly discriminative. With increase in target DNA/miRNA concentration, the difference in fluorescence intensity also increased, resulting in a good linear relationship between fluorescence intensity sensitizing value and target miRNA concentrations. Therefore, a new fluorescence analysis method for direct detection of let-7a in human gastric cancer cell samples without enzyme, label free and no immobilization was established using lucigenin/chitosan/SiO 2 composite nanoparticles as a DNA hybrid indicator. The proposed method had high sensitivity and selectivity, low cost and the detection limit was 10 fM (S/ N = 3).
研究领域:
纳米技术
·
生物传感器
·
癌症研究
·
分子生物学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
·
2021
·Vol. 19
·期 1
·1-18
·
IF 10.435
Wang Xueqin,Xiong Tiandi,Cui Miao,Li Na,Li Qin,Zhu Li,Duan Shaofeng,Wang Yunlong,Guo Yuqi
Background Multidrug resistance (MDR) is the main challenge of successful chemotherapy for ovarian cancer patients, with 50% to 75% of ovarian cancer patients eventually relapsed due to it. One of the effective strategies for treating MDR and improving therapeutic efficiency of ovarian cancer is to use nanotechnology-based targeted drug delivery systems. In this study, a novel nano targeted co-delivery system modified by hyaluronic acid (HA) was developed by using gold nanorods coated with functionalized mesoporous silica nanoparticles (HA-PTX/let-7a-GNR@MSN) for combined delivery of hydrophobic chemotherapy drug Paclitaxel (PTX) and lethal-7a (let-7a), a microRNA (miR), to overcome MDR in ovarian cancer. Furthermore, we also analyzed the molecular mechanism of this nanotherapeutic system in the treatment of ovarian cancer.Results HA-modified nanocomplexes can specifically bind to the CD44 receptor, which is highly expressed in SKOV3/SKOV3TR cells, achieving effective cell uptake and 150% enhancement of tumor site permeability. The nanosystem realized the stable combination and protective transportation of PTX and miRs. Analysis of drug-resistant SKOV3TR cells and an SKOV3TR xenograft model in BALB/c-nude mice showed significant downregulation of P-glycoprotein in heterogeneous tumor sites, PTX release, and subsequent induction of apoptosis. More importantly, this nanosystem could synergistically inhibit the growth of ovarian tumors. Further studies suggest that mTOR-mediated signaling pathways play an important role in reversing drug resistance and inducing apoptosis.Conclusion sTo sum up, these data provide a model for overcoming PTX resistance in ovarian cancer.Graphical Abstract
研究领域:
肿瘤学
·
纳米技术
·
药学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
ANALYST
·
2017
·Vol. 142
·期 15
·2796-2804
·
IF 3.885
Zhenqiang Wang,Jixi Zhang,Feng Chen,Kaiyong Cai
The combination of fluorophore-labelled single-strand DNA probes and nanomaterial quenchers has shown great potential in miRNA detection. The development of advanced detection systems by understanding and controlling the fluorescence quenching/recovery via nanoquenchers’ microstructures and local morphologies is an attractive area warranting further investigations. Inspired by nanopore sequencing, we present a novel miRNA sensing strategy using fluorophore-labeled DNA as probes and a type of large-pore-sized mesoporous polydopamine nanoparticles (MPDA-L, 70 nm in diameter) as fluorescence quenchers. It is revealed that the quenching efficiency of MPDA-L towards the fluorophore labelled on the probe, reached more than 99% at a relatively low particle concentration. Moreover, the mesopores effectively protected the probe DNA from cleavage by DNase I which was used for signal amplification. Sensitive detection of miRNA with a low detection limit of 32–40 pM, as well as a linear detection range of up to 5 nM, was realized by the mesopore effects via a greatly improved differential affinity of ssDNA and the probe–miRNA heteroduplex toward the surface of nanoquenchers. Interestingly, enhanced DLVO (Derjaguin–Landau–Verwey–Overbeek) repulsion generated inside the pore surface by the negative surface–curvature effect correlates with the improved duplex detachment and fluorescence recovery. The developed strategy can be successfully applied to quantify down-regulated let-7a and up-regulated miRNA-21 in different types of cancer cells by using total RNA samples from cell lysate. These findings are expected to inspire strategies and pave a way for utilizing porous nanomaterials for constructing miRNA detection systems.
研究领域:
纳米技术
·
生物医学工程
·
分子生物学
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）