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Chemical Science · 2020 ·Vol. 11 ·期 7 ·1948-1956 · IF 9.346

Capture and selective release of multiple types of circulating tumor cells using smart DNAzyme probes

Qianying Zhang,Wenjing Wang,Shan Huang,Sha Yu,Tingting Tan,Jian-Rong Zhang,Jun-Jie Zhu

The effective capture, release and reanalysis of circulating tumor cells (CTCs) are of great significance to acquire tumor information and promote the progress of tumor therapy. Particularly, the selective release of multiple types of CTCs is critical to further study; however, it is still a great challenge. To meet this challenge, we designed a smart DNAzyme probe-based platform. By combining multiple targeting aptamers and multiple metal ion responsive DNAzymes, efficient capture and selective release of multiple types CTCs were realized. Sgc8c aptamer integrated Cu2+-dependent DNAzyme and TD05 aptamer integrated Mg2+-dependent DNAzyme can capture CCRF-CEM cells and Ramos cells respectively on the substrate. With the addition of Cu2+ or Mg2+, CCRF-CEM cells or Ramos cells will be released from the substrate with specific selectivity. Furthermore, our platform has been successfully demonstrated in the whole blood sample. Therefore, our capture/release platform will benefit research on the molecular analysis of CTCs after release and has great potential for cancer diagnosis and individualized treatment.

研究领域: 生物技术 · 癌症研究 · 分子生物学
✓ 已验证 DOI PubMed 高影响因子
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Science China-Chemistry · 2019 ·Vol. 63 ·期 3 ·404-410 · IF 6.085

A pH-regulated stimuli-responsive strategy for RNA-cleaving DNAzyme

Lu Shasha,Wang Shuang,Zhao Jiahui,Sun Jian,Yang Xiurong

RNA-cleaving DNAzymes possess important roles in DNAzymes and have been widely used in the biosensors, DNA nanomachines owing to their ion-specific dependence. However, there are still challenges in constructing universal but versatile stimuli-responsive strategies of RNA-cleaving DNAzymes. Herein, a stimuli-responsive strategy for RNA-cleaving DNAzyme is proposed by the artful design of hairpin nanostructure, in which the activities of DNAzyme (Pb 2+ -dependent DNAzyme as a model) in the hairpin’s loop are pH-regulated by using the triplex stem as the “lock”. Upon introducing the “key”, pH values, the DNAzyme will be activated and fragment the substrate of it in the presence of Pb 2+ , accompanied by the turn-on of the fluorescence quenched by fluorescence resonance energy transfer (FRET). The regulation ability of pH can be controlled by the length and sequence of the triplex stem, and the wide pH regulation range may be helpful for the application of DNAzymes in biological medicine delivery systems.

CAS / MDL: 301-04-2
研究领域: 生物化学 · 纳米技术 · 生物传感器 · 分子生物学
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BIOELECTROCHEMISTRY · 2022 · IF 5.76

An electrochemical DNA sensor based on an integrated and automated DNA walker

Hao Fan,Ying Wu,Tongfu Huang,Nian Hong,Hanfeng Cui,Guobing Wei,Fusheng Liao,Jing Zhang

As an artificial nanomachine, a DNA walker demonstrates the potential for biosensing. In this study, a highly integrated, biostable, and autonomous electrochemical DNA walker sensor was rationally designed by a simple assembly of a Mn 2+ -dependent DNAzyme-powered DNA walker with nanoscale Mn 2+ @MOFs containing free carboxylic acid groups UiO-66(Zr)-(COOH) 2 . In this study, the release of Mn 2+ from Mn 2+ @MOFs was exploited to drive the autonomous and progressive operation of the DNA walker, and the DNAzyme-driven DNA walker was constructed by the co-modification of walking strands and track strands onto the gold electrode (GE) surface. The walking strand was a single-stranded DNA containing a DNAzyme sequence, which was pre-silenced by the locking strand. The track strand was a specially designed DNA sequence that the target can hybridize with the locking strand; hence, the walking strand is unlocked, and the liberated DNAzyme catalyzes the cleavage of track strands to drive the DNA walker operation, shifting tetraferrocene away from the electrode and producing a significant signal change. A detection limit of 38 fM was obtained with our new system, exhibiting a wide linear range from 1.5625 × 10 −9 M to 1 × 10 −13 M. The proposed approach provided a novel means for constructing an highly integrated, automated, and DNAzyme-driven DNA walker for bioanalysis.

研究领域: 生物传感 · 纳米技术 · 分子生物学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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TALANTA · 2014 · IF 3.511

G-quadruplex DNA biosensor for sensitive visible detection of genetically modified food

Xiaohua Jiang,Huimin Zhang,Jun Wu,Xiang Yang,Jingwei Shao,Yujing Lu,Bin Qiu,Zhenyu Lin,Guonan Chen

In this paper, a novel label-free G-quadruplex DNAzyme sensor has been proposed for colorimetric identification of GMO using CaMV 35S promoter sequence as the target. The binary probes can fold into G-quadruplex structure in the presence of DNA-T (Target DNA) and then combine with hemin to form a DNAzyme resembling horseradish peroxidase . The detection system consists of two G-rich probes with 2:2 split mode by using the absorbance and color of ABTS 2− as signal reporter. Upon the addition of a target sequence, two probes both hybridize with target and then their G-rich sequences combine to form a G-quadruplex DNAzyme, and the DNAzyme can catalyze the reaction of ABTS 2− with H 2 O 2 . Then the linear range is from 0.05 to 0.5   μM while detection limit is 5   nM. These results demonstrate that the proposed G-quadruplex DNAzyme method could be used as a simple, sensitive and cost-effective approach for assays of GMO.

研究领域: 生物化学 · 分子生物学 · 食品科学
✓ 已验证 DOI 高影响因子
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BIOSENSORS & BIOELECTRONICS · 2017 · IF 7.78

Target triggered cleavage effect of DNAzyme: Relying on Pd-Pt alloys functionalized Fe-MOFs for amplified detection of Pb2+

Yujie Yu,Chao Yu,Yazhen Niu,Jun Chen,Yilin Zhao,Yuchan Zhang,Rufei Gao,Junlin He

We designed an amplified detection strategy for the sensitive determination of lead ions (Pb 2+ ) based on a target-triggered nuclear acid cleavage of Pb 2+ -specific DNAzyme as a selectivity interface combined with Pd-Pt alloys modified Fe-MOFs (Fe-MOFs/PdPt NPs) hybrids acting as the signal tag. Streptavidin modified reduced graphene oxide-tetraethylene pentamine-gold nanoparticles (rGO-TEPA-Au) served as a sensor platform for immobilizing more DNAzyme. In the presence of Pb 2+ , the substrate DNA strand can be specifically cleaved at the ribonucleotide site by DNAzyme to produce a new single-DNA on the interface. Then, the hairpin DNA with hybrid strand matched by its complement to the single-DNA was employed to modify the Fe-MOFs/PdPt NPs bioconjugates for signal amplification . Fe-MOFs/PdPt NPs catalyze hydrogen peroxide (H 2 O 2 ) to produce the electrochemical signal which was recorded by chronoamperometry . Benefiting from the Pb 2+ -dependent DNAzyme, the proposed method can selectively detect Pb 2+ in the presence of other metal ions . The newly designed biosensor exhibited a good linear relationship ranging from 0.005 to 1000   nmol   L −1 with a low detection limit of 2   pM (S/N = 3) for Pb 2+ . This Pb 2+ -dependent DNAzyme based ultrasensitive biosensor showed high sensitivity and selectivity, providing potential application for Pb 2+ detection in naturally contaminated sewage and spiked drinking water samples.

研究领域: 生物化学 · 环境科学 · 分析化学
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ANALYTICAL AND BIOANALYTICAL CHEMISTRY · 2017 ·Vol. 409 ·期 27 ·6421-6427 · IF 3.431

Click chemistry-mediated cyclic cleavage of metal ion-dependent DNAzymes for amplified and colorimetric detection of human serum copper (II)

Li Daxiu,Xie Jiaqing,Zhou Wenjiao,Jiang Bingying,Yuan Ruo,Xiang Yun

The determination of the level of Cu 2+ plays important roles in disease diagnosis and environmental monitoring. By coupling Cu + -catalyzed click chemistry and metal ion-dependent DNAzyme cyclic amplification, we have developed a convenient and sensitive colorimetric sensing method for the detection of Cu 2+ in human serums. The target Cu 2+ can be reduced by ascorbate to form Cu + , which catalyzes the azide-alkyne cycloaddition between the azide- and alkyne-modified DNAs to form Mg 2+ -dependent DNAzymes. Subsequently, the Mg 2+ ions catalyze the cleavage of the hairpin DNA substrate sequences of the DNAzymes and trigger cyclic generation of a large number of free G-quadruplex sequences, which bind hemin to form the G-quadruplex/hemin artificial peroxidase to cause significant color transition of the sensing solution for sensitive colorimetric detection of Cu 2+ . This method shows a dynamic range of 5 to 500 nM and a detection limit of 2 nM for Cu 2+ detection. Besides, the level of Cu 2+ in human serums can also be determined by using this sensing approach. With the advantages of simplicity and high sensitivity, such sensing method thus holds great potential for on-site determination of Cu 2+ in different samples. Sensitive colorimetric detection of copper (II) by coupling click chemistry with metal ion-dependentDNAzymes

研究领域: 生物化学 · 分析化学
✓ 已验证 DOI PubMed 高影响因子
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Chemosensors · 2022 ·Vol. 10 ·期 7 ·276 · IF 4.229

Programmable, Universal DNAzyme Amplifier Supporting Pancreatic Cancer-Related miRNAs Detection

Kunhan Nie,Yongjian Jiang,Na Wang,Yajun Wang,Di Li,Lei Zhan,Chengzhi Huang,Chunmei Li

The abnormal expression of miRNA is closely related to the occurrence of pancreatic cancer. Herein, a programmable DNAzyme amplifier for the universal detection of pancreatic cancer-related miRNAs was proposed based on its programmability through the rational design of sequences. The fluorescence signal recovery of the DNAzyme amplifier showed a good linear relationship with the concentration of miR-10b in the range of 10–60 nM, with a detection limit of 893 pM. At the same time, this method displayed a high selectivity for miR-10b, with a remarkable discrimination of a single nucleotide difference. Furthermore, this method was also successfully used to detect miR-21 in the range of 10–60 nM based on the programmability of the DNA amplifier, exhibiting the universal application feasibility of this design. Overall, the proposed programmable DNAzyme cycle amplifier strategy shows promising potential for the simple, rapid, and universal detection of pancreatic cancer-related miRNAs, which is significant for improving the accuracy of pancreatic cancer diagnosis.Keywords:DNAzyme amplifier;signal amplification;miRNA detection;pancreatic cancer

CAS / MDL: 7773-01-5
研究领域: 分子生物学 · 医学诊断 · 生物化学
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FOOD CHEMISTRY · 2023 · IF 8.8

DNAzyme amplified dispersion state change of gold nanoparticles and its dual optical channels for ultrasensitive and facile detection of lead ion in preserved eggs

Wenju Yan,Xuefei Qin,Xueqing Sang,Xueying Zhou,Yanying Zheng,Yali Yuan,Yun Zhang

A dual-mode sensing platform for Pb 2+ was constructed based on the dual optical channels of Au NPs system with the amplification of DNAzyme, and it was successfully applied for Pb 2+ determination in preserved egg with satisfactory results. The presence of Pb 2+ activated the DNAzyme and induced the dispersion change of Au NPs in high salt concentration. The sequent absorption change of Au NPs was translated to the fluorescence change of carbon dots through FRET, and the scattering change was transferred to grey value of images involving the Tyndall effect. Thus, a sensing platform based on fluorescence and colorimetric dual-technique was achieved for Pb 2+ detection, under the optimized conditions. With the assistance of DNAzyme, the linear range of fluorometric and colorimetric method were 2.0 × 10 -14  ∼ 8.0 × 10 -10 mol/L and 2.4 × 10 -13  ∼ 9.5 × 10 -9 mol/L, respectively. The dual-mode sensing platform demonstrated its promising application in the environmental monitoring and food safety field.

CAS / MDL: 7447-39-4 10045-94-0
研究领域: 分析化学 · 环境科学 · 食品安全
✓ 已验证 产品关联 DOI PubMed 高影响因子
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BIOSENSORS & BIOELECTRONICS · 2013 · IF 5.437

Amplified colorimetric detection of mercuric ions through autonomous assembly of G-quadruplex DNAzyme nanowires

Yanli Hao,Qingquan Guo,Hanyin Wu,Liangqia Guo,Liangshuang Zhong,Jing Wang,Tianran Lin,FengFu Fu,Guonan Chen

An amplified colorimetric detection of Hg 2+ is proposed by combining T–Hg 2+ –T base pairs and hybridization chain reaction (HCR). Two hairpins consisting of three-fourths and one-fourth of the horseradish peroxidase (HRP)-mimicking DNAzyme in inactive configuration are used as functional elements. In the presence of Hg 2+ , one of the hairpins is opened by an assistant probe with the help of T–Hg 2+ –T base pairs and this triggers an autonomous cross-opening of the two hairpins using the strand displacement principle, resulting in the formation of DNA nanowires consisting of numerous reunited Q-quadruplex DNAzyme units. The resulting catalytically active hemin/G-quadruplex HRP-mimicking DNAzymes catalyze the oxidation of ABTS 2− by H 2 O 2 into a green-colored cationic radical ABTS •+ for the colorimetric readout. This “turn-on” sensing system enables the high sensitive and selective detection of aqueous Hg 2+ with a detection limit of 9.7   pM.

CAS / MDL: 16009-13-5
研究领域: 分析化学 · 纳米技术 · 环境科学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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MICROCHIMICA ACTA · 2022 ·Vol. 189 ·期 7 ·1-10 · IF 6.408

Dual DNAzyme-catalytic assembly of G-quadruplexes for inducing the aggregation of gold nanoparticles and developing a novel antibiotic assay method

Wang Xiaojun,Yang Jingru,Xie Yiming,Lai Guosong

By utilizing a target biorecognition reaction to induce the self-assembly of G-quadruplexes and the aggregation of gold nanoparticles (Au NPs), this work develops a novel colorimetric biosensing method for kanamycin (Kana) antibiotic detection. The compact G-quadruplex structure was assembled from its two half-split sequences which were designed in two hairpin substrates of the Mg 2+ -dependent DNAzyme (MNAzyme). Besides hybridizing with the aptamer strand, the MNAzyme sequence was also split into two half fragments to be designed in the two substrates. Upon the aptamer-recognition reaction toward Kana, the MNAzyme strand could be quantitatively released to cause the exposure of the split G-quadruplex-sequences on two hairpin substrate-modified Au NPs and simultaneous release of two half fragments of the MNAzyme-sequence. Thus, the K + -assisted self-folding of G-quadruplexes causes the cross-linking of the two Au NPs to realize the Au NP aggregation–based colorimetric signal output (measured at the largest absorption peak near 520 nm). Meanwhile, the self-assembled formation of the second MNAzyme drastically amplified the signal response. Under the optimal conditions, a wide linear range from 0.1 pg mL −1  to 10 ng mL −1 and an ultrahigh sensitivity with the detection limit of 76 fg mL −1 were obtained. The dose-recovery experiments in real samples showed satisfactory results with recoveries from 98.4 to 105.4% and relative errors compared with the ELISA method less than 4.1%. Due to the high selectivity, excellent repeatability and stability, and simple manipulation, this method indicates a promising potential for practical applications. Graphical abstract A novel homogeneous biosensing method was developed for the convenient detection of the kanamycin antibiotic. The target biorecognition-induced and dual DNAzyme-catalytic assembly of G-quadruplexes enabled the amplified aggregation of gold nanoparticles for the simple, cheap, stable, and ultrasensitive colorimetric signal transduction of the method.

研究领域: 分析化学 · 纳米技术 · 生物化学 · 生物传感器
✓ 已验证 产品关联 DOI PubMed 高影响因子
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TALANTA · 2021 · IF 6.057

Near-infrared light excited UCNP-DNAzyme nanosensor for selective detection of Pb2+ and in vivo imaging

Linna Huang,Feng Chen,Xia Zong,Qiujun Lu,Cuiyan Wu,Ziqi Ni,Meiling Liu,Youyu Zhang

Due to similar charges, atomic radii, and chemical properties of most metal ions , the selective sensing of these metal ions and imaging in vivo is still a challenge. A DNAzyme assemblied and near-infrared (NIR) light excited nanosensor was developed to detect and image Pb 2+ . In this nanosensor, NaYF 4 :Yb, Er upconversion nanoparticles (UCNPs) introduced as NIR-to-Vis transducer were the donor of luminescence resonance energy transfer (LRET), and DNAzyme-functionalized black hole quencher 1 (BHQ1) acted as energy transfer acceptor. This proposed nanosensor was applied to detecting Pb 2+ in solution with high sensitivity and selectivity . Furthermore, we have successfully demonstrated the imaging ability of this nanosensor towords Pb 2+ in living cells and early-stage zebrafish with negligible autofluorescence and good photostability. The UCNP-DNAzyme nanosensor would enrich the method of imaging Pb 2+ in vivo, and might serve as a potential tool for understanding the metabolic pathways of Pb 2+ and the mechanism of lead poisoning in biological system.

研究领域: 生物化学 · 分析化学 · 分子生物学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Science China-Chemistry · 2016 ·Vol. 60 ·期 2 ·293-298 · IF 2.429

Target-responsive DNAzyme hydrogel for portable colorimetric detection of lanthanide(III) ions

Huang Yishun,Wu Xuemeng,Tian Tian,Zhu Zhi,Lin Hui,Yang Chaoyong

Lanthanide elements (Ln) play an important role in industry and agriculture. As a result of the increasing consumption of lanthanides, environmental emission of Ln has become detrimental to the health of flora and fauna. Current methods for trace lanthanides detection mainly rely on sophisticated instruments. In this article, a Ln 3+ dependent DNAzyme was incorporated into a hydrogel to generate Ln 3+ sensitive DNAzyme hydrogel for portable colorimetric detection. The enzyme strand and its substrate strand act as crosslinker and functional unit of the hydrogel with polyacrylamide chains as the scaffold and gold nanoparticles (AuNPs) as the indicator of hydrogel stability. Any ions in the Ln 3+ series can trigger the cleavage of substrate strand by activating the enzyme strand, thereby decreasing the crosslink ratio and leading to collapse of the hydrogel. The release of the encapsulated AuNPs turns the supernatant wine red. Using this colorimetric method, Ln 3+ can be detected with high sensitivity, with a limit of detection (LOD) of 20 nM for Ce 3+ . The hydrogel responds specifically to any Ln 3+ ion and works well with the spiked lake sample without the need of instruments and skilled operators. Our results suggest that the lanthanide responsive hydrogel can be used for portable and sensitive detection of Ln 3+ contamination in the field.

研究领域: 生物化学 · 环境科学
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