ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Yuanyuan Yue,Songtao Wang,Qiang Jin,Nan An,Lina Wu,He Huang
Lead ions (Pb 2+ ) are a well-known toxic heavy metal that poses a significant threat to human health. Therefore, the development of a simple and ultrasensitive technique for detecting Pb 2+ is essential. With their trans -cleavage properties, the newly discovered CRISPR-V effectors have become a potential high-precision biometric tool. In this regard, a CRISPR/Cas12a-based electrochemical biosensor (E-CRISPR) has been developed, which is combined with the GR-5 DNAzyme that can specifically recognize Pb 2+ . In this strategy, the GR-5 DNAzyme acts as a signal-mediated intermediary, which can convert Pb 2+ into nucleic acid signals, thereby becoming single-stranded DNA that triggers strand displacement amplification (SDA) reaction. This is coupled with following activated CRISPR/Cas12a cleavage of the electrochemical signal probe, enabling cooperative signal amplification for ultrasensitive Pb 2+ detection. The proposed method has a detection limit as low as 0.02 pM. Therefore, we have developed an E-CRISPR detection platform with GR-5 DNAzyme as a signal medium (called SM-E-CRISPR biosensor). This provides a method for the CRISPR system to specifically detect non-nucleic substances by converting the signal using a medium.
研究领域:
生物化学
·
分子生物学
·
分析化学
·
环境科学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Bin Liu,Tian Wang,Dehui Qiu,Xinrong Yan,Yuan Liu,Jean-Louis Mergny,Xiaobo Zhang,David Monchaud,Huangxian Ju,Jun Zhou
Hemin/G-quadruplex (hG4) complexes are frequently used as artificial peroxidase-like enzymatic systems (termed G4 DNAzymes) in many biosensing applications, in spite of a rather low efficiency, notably in terms of detection limits. To tackle this issue, we report herein a strategy in which hemin is chemically modified with the amino acids found in the active site of parent horseradish peroxidase (HRP), with the aim of recreating an environment conducive to high catalytic activity. When hemin is conjugated with a single arginine, it associates with G4 to create an arginine–hemin/G4 (R-hG4) DNAzyme that exhibits improved catalytic performances, characterized by kinetic analysis and DFT calculations. The practical relevance of this system was demonstrated with the implementation of biosensing assays enabling the chemiluminescent detection of G4-containing DNA and colorimetry detection of the flap endonuclease 1 (FEN1) enzyme with a high efficiency and sensitivity. Our results thus provide a guide for future enzyme engineering campaigns to create ever more efficient peroxidase-mimicking DNA-based systems.
研究领域:
生物化学
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分子生物学
·
化学生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Yanli Lei,Xinyan He,Yu Zeng,Xinyu Wang,Le Yang,Xiaowen Liu,Zhihe Qing
Gold nanoparticles (Au NPs) have been widely utilized in developing DNAzyme-functionalized nanosensors, most of which were engineered by attaching the thiolated DNAzymes to Au NPs via Au-S bonding. However, the Au NP-DNAzyme nanosensors always suffer from signal distortion when applied in complex environment with abundant thiols, which poses challenge for practical applications. Here, we focus on addressing the root cause of the issue and propose to decorate the Au NPs with a thin layer of platinum, thus facilitating the conjugation of DNAzymes through Pt-S bonding, a thiol-resistant cross-linking. The Pt-S bond stabilized DNAzyme nanosensor effectively minimized false positive signals when detecting L-histidine in infant formulas, as compared to the Au-S stabilized counterpart. This innovative strategy holds promise for high-fidelity biosensing, improving the practical applicability of Au NP-based DNAzyme nanosensor.
研究领域:
生物化学
·
纳米生物技术
·
分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Shijiong Wei,Dehui Qiu,Xinrong Yan,Bin Liu,Jean-Louis Mergny,David Monchaud,Huangxian Ju,Jun Zhou
Colistin (COL) is a widely used antibiotic and is quite often used as a last-resort treatment option for treating multidrug-resistant Gram-negative bacterial infections. Due to its widespread use, COL accumulates in nature, which represents a novel ecological and health threat. However, there is currently no rapid and specific method available for titrating COL levels in collected samples. Herein, we report a simple chemiluminescence detection method based on the specific interaction between COL and a parallel G-quadruplex (G4). To this end, we exploit the catalytic properties of the G4/hemin DNAzyme, which is able to oxidize substrates to provide a readily monitored readout. The stronger affinity of G4 for COL versus hemin allows for the inactivation of the G4/hemin DNAzyme, which is used herein to quantify COL in solution. Through a series of optimizations, we identified the best G4 sequence (F3TC), oxidation substrate (luminol), and experimental conditions, which allow for the detection of COL over a broad concentration window, from 0.5 to 2,500 ng/mL, with a detection limit of 0.4 ng/mL and excellent selectivity against other antibiotics. Compared with existing methods, the proposed approach provides a simpler and label-free quantification of COL, which might serve as a valuable standard method for antibiotic detection, whose use was validated under real conditions herein.
研究领域:
分析化学
·
生物化学
·
抗生素检测
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环境监测
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF CONTROLLED RELEASE
·
2021
·
IF 9.776
Chao Wang,Zhaoyi Sun,Chenxuan Zhao,Zhewei Zhang,Haoran Wang,Yang Liu,Yunfei Guo,Bingtao Zhang,Lihong Gu,Yue Yu,Yiqiao Hu,Jinhui Wu
Radiotherapy (RT)-induced DNA damage leaked into cytosol can elicit host antitumor immune response. However, such response rate is unpromising due to limited cyclic GMP-AMP synthase (cGAS) recognition of cytosolic DNA, which could be digested inherently by host DNases . Here we show that synchronizing Mn 2+ delivery with accumulated cytosolic DNA after RT can promote the activation of cGAS-STING pathway, thereby enhancing RT-induced antitumor immunity. Intratumoral Mn 2+ injection immediately after RT cannot enhance RT, while intratumoral Mn 2+ injection 24 h after RT can. Direct-injected Mn 2+ can be metabolized out from tumor in minutes while RT-induced DNA damage need cells mitotic progression for up to 24 h to accumulate into cytosol. Alginate can maintain Mn 2+ in tumor for up to 24 h due to it can chelate divalent cations . When the release profile of Mn 2+ is controlled by alginate (Alg) and synchronized with the accumulation of RT-induced DNA damage, over 90% inhibition rate can be obtained even in the unirradiated tumor, and survival time is significantly extended. This synchronizing strategy provides a simple and novel approach to effectively activate cGAS-STING pathway in tumor and promote RT-induced immunity.
研究领域:
肿瘤学
·
免疫学
·
放射生物学
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DOI
PubMed
高影响因子
DALTON TRANSACTIONS
·
2020
·Vol. 49
·期 37
·13160-13166
·
IF 4.174
Yali Yu,Qingqing Zhang,Heng Gao,Chenxiao Yan,Xiong Zheng,Tong Yang,Xiaoshun Zhou,Yong Shao
G-quadruplex DNAs (G4s) have been reported to exhibit the DNAzyme activities by binding with some metal complexes and functional organic ligands. However, there is a challenge to develop metalloenzyme-mimic G4-based innate DNAzymes using the complexed metal ions directly serving as the active centers. This will diversify DNAzymes for developing novel devices since G4 structures are more polymorphic than the other DNA foldings. In this work, we found that the lanthanide trivalent cerium ion of Ce3+ can bind to the human telomere G4 (htG4) according to a 1 : 2 binding mode favorable for creating metalloenzymes-mimic G4 DNAzymes. This Ce3+-G4 entity exhibits a peroxidase activity towards the oxidation of the substrate of 3,3,5,5-tetramethylbenzidine (TMB) by hydrogen peroxide. The 5′ G4 tetrads with the orderly arranged carbonyl oxygen atoms are believed to be the coordination sites for Ce3+ and favor the conversion between Ce3+ and Ce4+. Our work provides an alternative feasibility in developing the G4-based innate DNAzymes for variant applications.
研究领域:
生物化学
·
分子生物学
·
催化
·
核酸化学
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DOI
PubMed
高影响因子
ANALYST
·
2019
·Vol. 144
·期 16
·4995-5002
·
IF 4.019
Rufeng Zhang,Yu Wang,Xiaonan Qu,Shasha Li,Yihan Zhao,Fenfen Zhang,Su Liu,Jiadong Huang,Jinghua Yu
Herein, a split G-quadruplex DNAzyme as a signal reporter was integrated into an electrochemical sensing platform for the detection of antibiotics with specificity and sensitivity. To improve the signal-to-noise ratio, two G-rich oligonucleotide sequences (G1 and G2) were blocked into two different hairpin probes, preventing the two segments from assembling into a spilt G-quadruplex structure. Moreover, we designed a double-arch probe, consisting of an aptamer as the recognition element and two-step enzymatic signal amplification. Concretely, the first is the Nt.BbvCI-assisted nicking cyclic reaction activated by target-aptamer binding, and the second is exonuclease III-aided cyclic amplification for generating abundant G1 and G2. The modified capture probe on the electrode was used to combine G1 and G2 to form the spilt G-quadruplex/hemin when K+ and hemin were present. This complex plays the role of DNAzyme with superior horseradish peroxidase activity in catalyzing the decomposition of H2O2. Under optimal conditions, this biosensor showed an excellent performance for sensing kanamycin with a detection limit of 83 fM for kanamycin concentrations ranging from 100 fM to 1 nM. Hence, the proposed strategy has potential as an efficient and actual platform for small molecule analysis.
研究领域:
生物化学
·
分析化学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
FOOD CONTROL
·
2024
·
IF 5.6
Jiong Liang,Yunzhe Zhang,Zikun Li,Xin Lu,Congyan Qi,Qian Yang,Wei Zhang
Aflatoxin B1 is a potent carcinogen widely found in agricultural products and feed. Therefore, investigating the development of convenient, sensitive, and precise techniques for detecting AFB1 is critical. This work proposes a ratiometric electrochemical aptasensor based on DNAzyme-driven tripedal DNA walker for enzyme-free AFB1 detection. The design of multiple legs reduces the moving time on the electrode surface and enhances the value of signal response. In this approach, the tagging of ferrocene and methylene blue as signal switch molecules diminishes the impact of interferences and significantly enhances the reproducibility (RSD: 2.825%). Meanwhile, the tripedal DNA walker and DNAzyme dual signal amplification enabled the sensor to detect AFB1 traces in cereals. We achieved trace detection of AFB1 with a detection limit of 61 fg/mL and a wide linear range from 0.0001 to 10 ng/mL. Furthermore, the aptasensor success rate was comparable to that of HPLC-PMP. The results demonstrate enormous potential for detection. Furthermore, the method can detect various targets by altering the target aptamer , displaying attractive scalability. This suggests its tremendous prospective in detecting mycotoxins in food in the future.
研究领域:
食品安全检测
·
生物传感技术
·
分子诊断
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产品关联
DOI
高影响因子
FOOD CHEMISTRY
·
2018
·
IF 4.946
Wan Huang,Hongyu Zhang,Guosong Lai,Shun Liu,Bo Li,Aimin Yu
By combination of the aptamer biorecognition with the colorimetric signal transduction of a DNAzyme-functionalized nanoprobe, a new biosensing method was developed for the rapid and sensitive detection of chloramphenicol (CAP). The nanoprobe was prepared through the functionalization of gold nanoparticles with the complementary oligonucleotide against aptamer and high-content hemin/G-quadruplex DNAzyme. When one-step incubating the nanoprobe and CAP at a constructed aptamer-magnetic bead (MB) biosensing platform, due to the competitive biorecognition reaction, the nanoprobes related with CAP amounts were quantitative captured onto the MB surface. Based on the catalytic reaction of the peroxidase-mimicking DNAzyme, a colored substance was produced for the colorimetric signal transduction of the method. Due to the great signal amplification of the nanoprobe, a very low detection limit down to 0.13 pg/mL was obtained. Considering the excellent performance of the aptasensing method and satisfactory results for milk sample experiments, it indicates good reliability for practical applications.
研究领域:
分析化学
·
生物传感器
·
食品安全
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
APPLIED SURFACE SCIENCE
·
2017
·
IF 3.387
Rujian Jiang,Zhirong Xin,Shiai Xu,Hengchong Shi,Huawei Yang,Lingjie Song,Shunjie Yan,Shifang Luan,Jinghua Yin,Ather Farooq Khan,Yonggang Li
Biomaterial-associated infections critically compromise the functionality and performance of the medical devices, and pose a serious threat to human healthcare. Recently, natural DNase enzyme has been recognized as a potent material to prevent bacterial adhesion and biofilm formation. However, the vulnerability of DNase dramatically limits its long-term performance in antibacterial applications. In this work, DNase-mimicking polymer brushes were constructed to mimic the DNA-cleavage activity as well as the macromolecular scaffold of the natural DNase. The bacteria repellent efficacy of DNase-mimicking polymer brush-functionalized surface was comparable to that of the DNase-functionalized surface. More importantly, due to their inherent stability, DNase-mimicking polymer brushes presented the much better performance in inhibiting bacterial biofilm development for prolonged periods of time, as compared to the natural DNase. The as-developed DNase-mimicking polymer brush-functionalized surface presents a promising approach to combat biomaterial-associated infections.
研究领域:
生物医学工程
·
材料科学
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Li Wang,Wanwan Wang,Shumin Zhang,Jie Wei,Qingmin Chen,Tianhui Jiao,Anhui Lin,Quansheng Chen,Xiao-Mei Chen
In this study, we present an intelligent electromagnetic-actuated microfluidic chip integrated with a G-quadruplex DNAzyme-based biocatalysis platform for rapid and sensitive tetracycline (TC) detection. In this sensing system, TC significantly quenches fluorescent magnetic carbon dots (M-CDs) via the internal filtration effect and dynamic quenching (the excitation and emission wavelength at 350 and 440 nm, respectively). Then, the G-quadruplex on the M-CDs-Aptamer is exposed and bound with hemin to form hemin-G-quadruplex DNAzyme, catalyzing the conversion of 3,3′,5,5′-tetramethylbenzidine to produce blue color. This enables the fluorescence/colorimetric detection of TC. Importantly, an automatic electromagnet-integrated microfluidic chip was designed to control the shuttling of magnetic materials in each function slot according to a programmed sequence. Under the optimal conditions, the detection limits of TC for fluorescence and colorimetric methods were 11 and 43 μmol/L, respectively. The detection results for tilapia (Oreochromis nilotica) were comparable to those of traditional high-performance liquid chromatography. This platform offers excellent performance for TC determination and potential for portable, intelligent detection of trace pollutants in food and the environment.
研究领域:
生物化学
·
环境科学
·
食品科学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Xiaoxia Liu,Fang Yang,Daxiu Li,Ruo Yuan,Yun Xiang
Highly sensitive detection of protein biomarkers has an important role in clinical diagnosis and disease treatment. On the basis of a newly designed ternary probe containing the aptamer, primer and circular DNA sequences, we have established an ultrasensitive and label-free colorimetric approach for detecting platelet-derived growth factor BB protein biomarkers via a target-triggered autonomous multi-branch rolling circle amplification (mbRCA) strategy. The target molecules specifically bind the aptamer sequences in the ternary probes and causes the configuration transition of the primers to trigger mbRCA generation of numerous of G-quadruplex sequences with the presence of the assistance hairpin. Hemin further associates with these G-quadruplexes to yield many hemin/G-quadruplex DNAzymes with the horseradish peroxidase-mimicking function, which catalyze the substrate solution to induce intense color transition for ultrasensitive and label-free colorimetric determination of the target molecules with a 0.055 pg mL −1 of detection limit in the range of 0.1–1000 pg mL −1 . This aptamer-based sensing method exhibits high selectivity and can achieve the detection of low levels of protein biomarkers in diluted serum samples, which offers the developed approach new opportunities for detecting other trace protein biomarkers for the purpose of early disease diagnosis.
研究领域:
生物化学
·
分子生物学
·
医学诊断
✓ 已验证
DOI
高影响因子
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