TALANTA
·
2021
·
IF 6.057
Ting Huang,Guixun Chen,Birong Liu,Zizhong Yang,Yuanwei Huang,Baoping Xie,Min-Min Li,Jin-Xiang Chen,Jun Chen,Zong Dai
Sensitive, specific and rapid methods for detecting microRNAs (miRNAs) play critical roles in disease diagnosis and therapy. Enzyme-free amplification techniques based on DNAzyme assembly have recently been developed for the highly specific miRNA analysis. However, traditional DNAzyme-based assembly (free DNAzyme) amplifiers is mainly dependent on the target-induced split DNAzyme fragments to assemble into activated DNAzyme structures, which have made a compromise between the sensitivity and specificity due to the random diffusion of dissociative probes in a bulk solution with poor kinetics. Herein, based on a rationally designed DNA probe , we developed an intramolecular DNAzyme assembly (intra-DNAzyme) method to overcome these challenges. The miR-373 is used as model analyte for our current proof-of-concept experiments. Compared with the free-DNAzyme method, our method showed significantly improved analytical performance in terms of dynamic range, assay sensitivity and speed. This method can detect miR-373 specifically with a detection limit as low as 4.3 fM, which is about 83.7 times lower than the previous free-DNAzyme method. This intra-DNAzyme strategy would be of great value in both basic research and clinical diagnosis.
研究领域:
分子生物学
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生物技术
·
纳米技术
·
诊断学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Yong-Qiang He,Yu Gao,Hui-Wen Gu,Xian-Zhu Meng,Hong-Chao Yi,Ying Chen,Wei-Yin Sun
In this work, a sensitive electrochemical method for bleomycin (BLM) determination was reported on the basis of BLM-mediated activation of Zn 2+ -dependent DNAzyme and the adsorption of signal probes by a metal-organic framework (MOF) modified electrode . Two hairpin DNAs were employed in this protocol, one (HP1) for BLM recognition and one (HP2) for amplified signal output. The presence of BLM and Fe 2+ caused the formation of BLM-Fe (II) complex to cleave HP1, releasing DNAzyme fragments, which could further hybridize with substrate HP2 to form a partial double-stranded DNA duplex and enable the activation of Zn 2+ -dependent DNAzyme with the coexistence of Zn 2+ . The Zn 2+ -dependent DNAzyme catalyzed the cyclic cleavage of magnetic beads (MB)-immobilized HP2 to release massive DNA fragments with a Fc-labeled- terminal, which could be used for BLM quantification through electrochemical measurement after their adsorption on a MOF modified electrode. Attributing to the high catalytic efficiency of DNAzyme and excellent electrochemical performance of MOF modified electrode, our method revealed an impressive limit of detection as low as 4 pM BLM with a linear range of 5–2000 pM. Besides, the easy synthesis of MOF without further modification and the easy way of adsorption for signal achievement facilitated the operation process. In virtue of the high sensitivity, selectivity and the simple-to-implement features, this method is believed to hold a great promising application for BLM determination in biomedical and clinical study.
研究领域:
分析化学
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生物化学
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生物医学工程
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF ENVIRONMENTAL ANALYTICAL CHEMISTRY
·
2014
·
IF 1.321
Dandan Ma,Yali Yuan,Xilin Xiao,Qiong Tang,Yangyang Gao
A novel highly sensitive and specific electrochemical biosensor for detecting uranium based on specific Deoxyribozymes and gold nanoparticles (AuNPs) is reported. In this work, AuNPs provide excellent electrochemical signal transduction and a large surface area for immobilising numerous Deoxyribozymes, so a low detection limit of 3.24 ng L−1 uranium and a good linear relationship over the range 5.94–35.1 ng L−1 (r = 0.994) were obtained. The proposed biosensor presents high specificity and selectivity for uranium and is not affected by other metal ions. Thus, the biosensor protocol offers good selectivity, rapid speed and operational convenience for detection uranium in liquid waste.
研究领域:
生物化学
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环境科学
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2024
·
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
·
诊断方法
✓ 已验证
DOI
PubMed
高影响因子
ACS Nano
·
2018
·
IF 13.709
Jie Feng,Zhen Xu,Feng Liu,Yun Zhao,Wenqian Yu,Min Pan,Fuan Wang,Xiaoqing Liu
Catalytic deoxyribozyme has great potential for gene regulation, but the poor efficiency of the cleavage of mRNA and the lack of versatile DNAzyme vehicles remain big challenges for potent gene therapy. By the rational designing of a diverse vehicle of polydopamine-Mn2+ nanoparticles (MnPDA), we demonstrate that MnPDA has integrated functions as an effective DNAzyme delivery vector, a self-generation source of DNAzyme cofactor for catalytic mRNA cleavage, and an inherent therapeutic photothermal agent as well as contrast agent for photoacoustic and magnetic resonance imaging. Specifically, the DNAzyme-MnPDA nanosystem protects catalytic deoxyribozyme from degradation and enhances cellular uptake efficiency. In the presence of intracellular glutathione, the nanoparticles are able to in situ generate free Mn2+ as a cofactor of DNAzyme to effectively trigger the catalytic cleavage of mRNA for gene silencing. In addition, the nanosystem shows high photothermal conversion efficiency and excellent stability against photothermal processing and degradation in complex environments. Unlike previous DNAzyme delivery vehicles, this vehicle exhibits diverse functionalities for potent gene regulation, allowing multimodal imaging-guided synergetic gene regulation and photothermal therapy both in vitro and in vivo.
研究领域:
生物化学
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分子生物学
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诊疗学
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纳米医学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BULLETIN OF THE KOREAN CHEMICAL SOCIETY
·
2017
·Vol. 38
·期 10
·1143-1148
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IF 0.602
Mingjing Zou,Shuyu Wang
We report a visualization assay for carcinoembryonic antigen (CEA). The assay was prepared based on target-specific recognition, aptamer-based DNAzyme self-catalysis, and colorimetric reaction. Aptamer of CEA was gathered at the surface of magnetic beads for sandwiched co-conjugate when CEA was added. The second aptamer (Apt2) on the beads was folded into G-quadruplex by KCl to make G-quadruplex-hemin DNAzyme with hemin. Apt2-based DNAzyme catalyzed H 2 O 2 -mediated oxidation of 3,3′,5,5′-tetramethylbenzidine, resulting in color changes and significant increase in absorption intensity at 450 nm (Δ A 450nm ). Yellow products can be observed with naked eyes when over 1.0 ng/mL CEA was added and there was a linear correlation between Δ A 450 nm and CEA concentrations in the range of 0.1–8.0 ng/mL with the detection limit of 0.06 ng/mL. The proposed method was then used to determine CEA in human serum from healthy people and tumor patients, revealing a correlation with data from the electrochemiluminescent immunoassay. Owing to its simplicity, selectivity, and low-cost, this method is a promising candidate for detecting CEA and clinical diagnosis of cancer.
研究领域:
生物化学
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分析化学
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分子生物学
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临床诊断
✓ 已验证
产品关联
DOI
高影响因子
Chemosensors
·
2022
·Vol. 10
·期 12
·495
·
IF 4.229
Jingyi Si,Wei Zhou,Da Zhou,Ying Fang,Xizhong Shen,Changfeng Zhu
Calprotectin (CP) is an established biomarker that allows the noninvasive evaluation of inflammation levels in the gastrointestinal tract of patients with inflammatory bowel disease and is helpful for the diagnosis and management of the disease. Herein, we demonstrate that CP can effectively suppress the activity of 17E DNAzyme (17E) by chelating Zn(Ⅱ), which is the cofactor of 17E. As the inhibition efficiency of CP on the DNAzyme is proportional to the concentration of CP, the detection of CP can be readily achieved by assessing the activity of 17E.Keywords:DNAzyme;calprotectin;inflammatory bowel disease;fluorescent sensor;metal ion
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
✓ 已验证
产品关联
DOI
高影响因子
ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Jian Chu,Chun Chen,Xijian Li,Libing Yu,Wenjing Li,Mengxi Cheng,Wei Tang,Zhonghua Xiong
It is of great significance to develop facile and economical strategies for on-site detection and treatment of toxic metal ions . Stimulus-responsive DNA hydrogel materials have been increasingly used for convenient detection of metal ions due to their advantages such as simplicity, portability, and ease of storage. However, these methods still require encapsulation of signal tags by labeling or embedding. In this paper, a one-step preparation of Pb 2+ -responsive pure DNA hydrogel material was designed to realize a new label-free strategy for Pb 2+ biosensing. The Pb 2+ -dependent DNAzyme strand and substrate strand were introduced to fabricate the DNA hydrogel. The presence of Pb 2+ in the sample activates the enzyme strand in the hydrogel skeleton and triggers the cleavage of the substrate, thereby destroy the hydrogel structure. DNA fragments released by the collapsed hydrogel were readily measured as signal output for quantifying Pb 2+ concentrations with a minimum detection limit of 7.7 nM. We successfully eliminated the need for embedding or labeling of signal molecules by using the DNA molecules that construct hydrogels as the signal output. And the newly developed method for label-free detection of Pb 2+ based on pure DNA hydrogel is simple, easy readout, and cost-effective. By adjusting the DNAzyme and substrate sequences, label-free analysis of other metal ions can also be achieved. We expect that our strategy can be applied to the field detection of toxic metal ions.
研究领域:
环境科学
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材料科学
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生物传感
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2022
·Vol. 12
·期 29
·18296-18300
·
IF 4.036
Xiaofei Liao,Mengyan Li,Li Zou
In this work, a label-free and sensitive colorimetric sensing strategy for the detection of bleomycin (BLM) was developed on the basis of BLM-mediated activation of G-quadruplex DNAzyme via DNA strand scission. A G-quadruplex based hairpin probe (G4HP) containing the scission site (5′-GT-3′) of BLM at the loop region and guanine (G)-rich sequences at its 5′-end was employed in this protocol. In the presence of BLM, it may cleave the 5′-GT-3′ site of the hairpin probe with Fe(II) as a cofactor, releasing the G-tetrads DNA fragment, which may further bind hemin to form a catalytic G-quadruplex-hemin DNAzyme. The resultant G-quadruplex DNAzyme has notable peroxidase-like activity, which effectively catalyzes the oxidation of 2,2′-azino-bis(3-ethylbenzothiozoline-6-sulfonic acid) (ABTS) by H2O2 to produce the blue-green-colored free-radical cation (ABTS·+). Therefore, the detection of BLM can be achieved by observing the color transition with the naked eye or measuring the absorbance at a wavelength of 420 nm using a UV-Vis spectrophotometer. Attributing to the specific BLM-induced DNA strand scission and the effective locking of G-tetrads in the stem of the G4HP, the colorimetric sensing strategy exhibits high sensitivity and selectivity for detection of BLM in human serum samples, which might hold great promise for BLM assay in biomedical and clinical research.
研究领域:
生物医学工程
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分析化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 12
·1-8
·
IF 6.232
Yang Zongqi,Liu Mei,Li Baoxin
A DNAzyme-powered DNA walking machine was constructed to develop the fluorescence aptasensing for sensitive detection of kanamycin. The aptamer for kanamycin is partially hybridized with complementary DNA (cDNA) modified on magnetic beads (MBs). The specific interaction of target and aptamer triggered the cDNA to be free tentatively, which captured walker DNA. Then the autonomous motion of DNA walker on MBs surface was propelled via DNAzyme digestion of recognition sites. The signal probe was separated, and the amplified fluorescence signal was achieved by the accumulation of the signal probe. Kanamycin was used as a model analyte, and the developed assay achieves a detection limit of 0.00039 ng·mL −1 (S/ N = 3) within a linear detection range from 0.001 to 2000 ng·mL −1 . This aptasensing strategy can be extended for detection of other antibiotics by adapting corresponding target recognition aptamer sequence.
Graphical abstract The fluorescence aptasensing for sensitive detection of kanamycin based on DNAzyme-powered DNA walking machine was constructed.
研究领域:
分析化学
·
生物传感器
·
分子诊断
✓ 已验证
DOI
PubMed
高影响因子
ANALYST
·
2019
·Vol. 144
·期 17
·5098-5107
·
IF 4.019
Shengfeng Wang,Jinsong Ding,Wenhu Zhou
Simultaneous detection and regulation of tumor-related genes presents a promising strategy for early diagnosis and treatment of cancer, but achieving this has been a huge challenge for both chemical and biomedical communities. Towards this objective, we have devised a novel aptamer-tethered, DNAzyme-embedded molecular beacon (MB) for multiple functions in cancer cells. In this design, a tumor targeting aptamer was employed to specifically deliver the sensor into cancer cells for target gene detection, and an RNA-cleaving DNAzyme was embedded to realize gene regulation. Both aptamer-tethering and DNAzyme-embedding had little influence on the sensor performance, with a detection limit of ∼2 nM and high specificity. After delivering into tumor cells, our device could monitor the tumor-related genes by producing detectable fluorescence signals, and regulate the gene expression at both mRNA and protein levels as evidenced by the RT-PCR and western blot analyses. This study provides a simple and efficient strategy to rationally combine various functional nucleic acids for multi-functional applications in living cells, which hold great potential for cancer diagnosis and therapy.
研究领域:
生物技术
·
分子生物学
·
癌症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2015
·Vol. 3
·期 24
·4805-4813
·
IF 4.726
Rong Tian,Xiaojun Chen,Nan Jiang,Ning Hao,Lin Xu,Cheng Yao
A highly sensitive electrochemical sensor for the detection of mercury(II) ions (Hg2+) was developed by using a three dimensional ordered macroporous polyaniline–platinum (3DOM PANI–Pt) composite film as a sensing platform. 3DOM PANI–Pt was realized by electrochemical co-deposition of Pt nanoparticles and PANI into the sacrificed silica template through the redox reaction involving the monomer aniline (AN) and PtCl62−. The G-rich oligonucleotide strand functionalized Au nanoparticle was employed as the Hg2+-mediated structural switch and to enhance the sensitivity. In the absence of Hg2+, the oligonucleotide strand formed an intramolecular duplex where the G-rich sequence was partially caged. In the presence of Hg2+, the release of the G-rich sequence was observed due to the stabilization of T–Hg2+–T, which formed an active G-quadruplex DNAzyme and catalyzed the reduction of H2O2. Under the optimal conditions, the current signal of H2O2 increased with increasing Hg2+ concentration in the dynamic range from 10−13 to 10−6 M, and the detection limit up to 8.7 × 10−14 M was seen. Further, the sensor was successfully utilized for the determination of Hg2+ in an authentic aquatic sample, with an acceptable accuracy compared to the method commercially available. In this paper, we proposed a general and simple strategy because only one oligonucleotide strand was required for both Hg2+ recognition and signal amplification, potentially allowing the detection of other metal ions or trace pollutants in environmental matrices by simply employing various DNA or aptamer probes.
研究领域:
分析化学
·
环境科学
·
纳米技术
·
生物化学
✓ 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）