Frontiers in Bioengineering and Biotechnology
·
2022
·
IF 6.064
Li Yali,Wang Jing,Huang Fenghong,Zhang Yufei,Zheng Mingming
The artificial multienzyme systems developed by mimicking nature has attracted much interest. However, precisely controlled compositions and ratios of multienzymatic co-immobilization systems are still limited by the indistinguishable nature of enzymes. Herein, a strategy for fabricating DNA-directed immobilization of horseradish peroxidase (HRP) and glucose oxidase (GOx) on hybrid DNA nanoflowers (GOx-HRP@hDFs) is presented. The preparation of micron-sized hybrid DNA flowers (hDFs) begins with the predetermined repeatable polymer-like DNA sequences which contained two strands. The hDFs structure is generated through one-pot rolling circle amplification (RCA) and self-assembly with magnesium pyrophosphate inorganic crystals. Based on the rigid-base pairing, GOx and HRP conjugated with sequences complementary to strands would be anchored to the predesigned locations, respectively. By adjusting the loading amount/ratio of enzymes properly, the maximal catalytic efficiency can be precisely regulated. The reaction activity of GOx-HRP@hDFs was 7.4 times higher than that of the free GOx-HRP under the optimal mole ratio (GOx/HRP 4:1). In addition, this multienzyme catalyst system exhibits excellent precision, specificity, reproducibility, and long-term storage stability when applied to real human blood samples. The preceding results validate that GOx-HRP@hDFs are promising candidates for personal diabetes detection.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Water Biology and Security
·
2023
·
IF 0
Chaolin Jiang,Xingchen Huo,Lingjie Tang,Meidi Hu,Chunrong Yang,Daji Luo,Jianguo Su
Grass carp hemorrhagic disease caused by grass carp reovirus (GCRV) results in significant economic losses to the global grass carp aquaculture industry. Oral vaccination is an ideal choice for disease precaution in aquaculture. However, oral vaccine can be degraded in the gut. Therefore, the selection of loading materials is essential. In this study, the S6 and S7 fragments (encoding the outer capsid protein VP4 and fibronectin VP56 of GCRV) and grass carp interferons (IFNs), including IFN1, IFN3, and IFNγ2 were used to create DNA vaccines and adjuvants based on pcDNA3.1, respectively. The oral DNA vaccine was encapsulated in poly(lactic-co-glycolic acid) (PLGA) and polyvinyl alcohol (PVA) with IFNs. The PLGA-PVA (PP) nano-microspheres were prepared by double emulsion-solvent evaporation technique. Using transmission electron microscopy and dynamic light scattering assays, it was determined the vaccines had a spherical structure with uniform particle size (643.5 ± 35.3 nm). The nano-microspheres possessed excellent encapsulation efficiency (81.6 ± 2.6%) and loading rate (0.54 ± 0.02%), and simultaneously exhibited negligible hemolytic activity and cell toxicity. The protection rate and tissue viral loads post-GCRV challenge in grass carp were assessed. The oral PP nano-microsphere with pVP4 + pIFN1 (PP41) vaccine increased protection rate by 44% compared with the control group and was correlated with relatively low viral loads in the spleen, head kidney, and hindgut. Further, three crucial serum biochemical indexes, total superoxide dismutase (TSOD), complement C3 (C3), and lysozyme (LZM), were also dramatically increased. Furthermore, mRNA expressions of representative immune-related genes (IgM, IFN1, IFNγ2, MHC-Ⅰ, and CD8α) in the head kidney and spleen were significantly enhanced. In addition, mRNA expression of IgT was significantly boosted in the hindgut. The results indicate that DNA vaccine capsulated with PP is effective against GCRV infection. The present study provides insights into a prospective strategy for oral vaccine development in aquaculture.
研究领域:
水产养殖
·
免疫学
·
病毒学
·
兽医学
✓ 已验证
DOI
ACS Applied Materials & Interfaces
·
2020
·
IF 8.758
Zhenqiang Fan,Zongqiong Lin,Zepeng Wang,Jianfeng Wang,Minhao Xie,Jianfeng Zhao,Kai Zhang,Wei Huang
In this work, we fabricated a dual-wavelength electrochemiluminescence ratiometric biosensor based on electrochemiluminescent resonance energy transfer (ECL-RET). In this biosensor, Au nanoparticle-loaded graphitic phase carbon nitride (Au-g-C3N4) as a donor and Au-modified dimethylthiodiaminoterephthalate (TAT) analogue (Au@TAT) as an acceptor were investigated for the first time. Besides, tetrahedron DNA probe was immobilized onto Au-g-C3N4 to improve the binding efficiency of the transcription factor and ECL ratiometric changes on the basis of the ratio of ECL intensities at 595 and 460 nm, which were obtained through the formation of a sandwich structure of DNA probe–antigen–antibody. Our biosensor achieved the assay of NF-κB p50 with a detection limit of 5.8 pM as well as high stability and specificity.
研究领域:
分析化学
·
生物传感器
·
电化学
·
纳米技术
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Huijie Bai,Xue Wu,Yuanyi Zhao,Juan Hu,Junda Wang,Ziming Ye,Cuiping Mao,Yi Li,Kuo Chen,Yong Wang,Jinhong Guo
The development of a spatiotemporally resolved analytical platform capable of real-time, simultaneous tracking of base excision repair (BER) pathway-associated enzymes in living cellular systems holds significant potential for elucidating their cooperative mechanisms in disease pathogenesis and acquired therapeutic resistance. However, current simultaneous enzymatic imaging is constrained by probe integration complexity, lack inherent signal amplification capacity, and diffusion-dominated kinetics compromising response time, catalytic efficiency, and sensitivity. Herein, we report a magnetic nanomotor-enhanced double confined hybridization chain reaction (MEDC-HCR) to detect the correlated activity of apurinic/apyrimidinic endonuclease 1 (APE1) and flap structure-specific endonuclease 1 (FEN1) in an AND-gated manner. The hairpin-structured of the hybridization chain reaction (HCR) integrate dual enzymatic sensing modules for APE1 and FEN1 detection. Furthermore, the magnetic nanomotors exhibit dual functionality as confined scaffolds and efficient intracellular delivery vehicles, synergistically enhancing local probe concentration to accelerate reaction kinetics, and simultaneously improving probe stability in complex biological fluids. Clinical validation demonstrating that this method distinguishes between healthy individuals and breast cancer patients confirms its potential for clinical translation. The innovative platform serves as a versatile tool for investigating DNA repair mechanisms relevant to cancer diagnosis and treatment, and simultaneously highlights the potential of self-propelled nanomotors for molecular imaging and disease diagnostics.
✓ 已验证
产品关联
DOI
高影响因子
International Journal of Electrochemical Science
·
2023
·
IF 1.5
Mingyi Ma,Li He,Xiaoxue Shi,Yanchao Wang,Hong Hai,Xiaoping Wei
This paper presents a novel strategy for ultrasensitive detection of human immunodeficiency virus (HIV) DNA by an electrochemical biosensor. By the shearing cycle of the target DNA (T-DNA), a large amount of complimentary sequence probes (CS probe DNA) are obtained. Then, the Zr 4+ functional probe DNA containing many branched DNAs is captured. Next, the initiator α-bromophenylacetic acid is connected to the branched chains of probe DNA by esterification reaction. Numerous electroactive monomers, namely, ferrocenylmethyl methacrylate (FMMA), are produced at the end of the branched chains by electrochemically mediated atomic transfer radical polymerization reaction. The signal of FMMA is greatly enhanced due to a large number of branched chain-loaded probe clusters containing numerous FMMA. T-DNA can be detected with a linear range of 0.1 fM–10 nM with a detection limit of 0.037 fM. The sensor is highly sensitive and selective. It has been applied to the detection of HIV DNA in human serum samples, and it has potential application prospects in the early clinical diagnosis of AIDS.
✓ 已验证
产品关联
DOI
高影响因子
VIROLOGY
·
2023
·
IF 3.7
Xiaoping Huang,Xiao Wang,Yaqi Sun,Ling Li,Anna Li,Wentao Xu,Xiaolan Xie,Yong Diao
Because it is safe and has a simple genome, recombinant adeno-associated virus (rAAV) is an extremely appealing vector for delivery in in vivo gene therapy. However, its low transduction efficiency for some cells, limits its further application in the field of gene therapy. Bleomycin is a chemotherapeutic agent approved by the FDA whose effect on rAAV transduction has not been studied. In this study, we systematically investigated the effect of Bleomycin on the second-strand synthesis and used CRISPR/CAS9 and RNAi methods to understand the effects of Bleomycin on rAAV vector transduction, particularly the effect of DNA repair enzymes. The results showed that Bleomycin could promote rAAV2 transduction both in vivo and in vitro. Increased transduction was discovered to be a direct result of decreased cytoplasmic rAAV particle degradation and increased second-strand synthesis. TDP1 , PNKP , and SETMAR are required to repair the DNA damage gap caused by Bleomycin, TDP1, PNKP, and SETMAR promote rAAV second-strand synthesis. Bleomycin induced DNA-PKcs phosphorylation and phosphorylated DNA-PKcs and Artemis promoted second-strand synthesis. The current study identifies an effective method for increasing the capability and scope of in-vivo and in-vitro rAAV applications, which can amplify cell transduction at Bleomycin concentrations. It also supplies information on combining tumor gene therapy with chemotherapy.
✓ 已验证
DOI
PubMed
高影响因子
Nanoscale
·
2018
·Vol. 10
·期 12
·5532-5538
·
IF 7.233
Bi-Yun Fang,Cheng Li,Jie An,Sun-Duo Zhao,Zhi-Yong Zhuang,Yuan-Di Zhao,Yue-Xuan Zhang
In this paper, DNA containing six cytosines as the formation site for silver nanoclusters (Ag NCs) was adopted as a template for preparing fluorescent DNA-Ag NCs. For the first time, it was found that the fluorescence of DNA-Ag NCs could be quenched after hybridization with their complementary sequence. On the basis of this new phenomenon, we designed a sequence C1 that was completely complementary to human immunodeficiency virus (HIV) DNA, and probe DNA which was partially complementary to C1 for the synthesis of DNA-Ag NCs. The fluorescence of DNA-Ag NCs was quenched after hybridization with C1 and the DNA-Ag NCs/C1 composite was formed, while C1 could be dissociated away from the DNA-Ag NCs by HIV DNA through a strand exchange reaction due to the stronger affinity between HIV DNA and C1, which could switch on the quenched Ag NCs, thus a new “off–on” fluorescence method for HIV DNA detection was developed. In the experiment, the Ag NCs formation site of DNA, the number of base pairs, and the pH and salt concentration of binding buffer were optimized. Under the optimum conditions, the limit of detection for HIV DNA was obtained to be 3.18 nM (3σ/N, n = 7) with the linear range of 15–150 nM for the 150 nM DNA-Ag NCs/C1 probe. Besides, the probe showed excellent specificity to HIV DNA, and even distinguished one nucleotide mismatched HIV DNA.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ChemistrySelect
·
2022
·Vol. 7
·期 35
·e202202565
·
IF 2.307
Chunying Wei,Haiying Li
Graphical The compounds can bind selectively to G-quadruplex DNA and result in a strong turn-on red fluorescence response, and it is also a potential fluorescence probe for G-quadruplex DNA in the living cells. The small molecule fluorescence probes for G-quadruplexes (G4) play a vital role in the detection of G4. In this work, two novel D−π−A benzothiazole derivatives were synthesized, and their interactions with different secondary structure DNA, cytotoxicity, and intracellular distribution were investigated in detail. Both compounds have a high affinity and about 100-fold turn-on fluorescence response to the promoter G4 in buffer solution, moreover, the compound also stabilizes effectively the promoter G4 structure. Besides, both compounds have no influence on the conformation of the promoter G4, but they induce the folding of bcl-2 from the coiled chain into the mixed-type G4 in salt-free buffer solution. The binding stoichiometries of the compound to bcl-2, c-kit2, and c-myc G4 are near 2 : 1, 1 : 1, and 3 : 1, respectively, and the interactions are an enthalpy-driven process. Furthermore, both compounds inhibit the proliferation of HepG2 cells, with IC 50 values of 3.23 and 38.9 μM. The compound mainly distributes in the cytoplasm of living cells and might interact with DNA. The current results provide the valuable information for design of fluorescence probes targeting G4 in the living cell.
✓ 已验证
产品关联
DOI
高影响因子
Biosensors-Basel
·
2024
·Vol. 14
·期 2
·105
·
IF 4.9
Xinmeng Zhang,Cheng Zhou,Jianxun Hou,Gang Feng,Zhourui Xu,Yonghong Shao,Chengbin Yang,Gaixia Xu
Precise DNA quantification and nuclear imaging are pivotal for clinical testing, pathological diagnosis, and drug development. The detection and localization of mitochondrial DNA serve as crucial indicators of cellular health. We introduce a novel conjugated oligoelectrolyte (COE) molecule, COE-S3, featuring a planar backbone composed of three benzene rings and terminal side chains. This unique amphiphilic structure endows COE-S3 with exceptional water solubility, a high quantum yield of 0.79, and a significant fluorescence Stokes shift (λex= 366 nm, λem= 476 nm), alongside a specific fluorescence response to DNA. The fluorescence intensity correlates proportionally with DNA concentration. COE-S3 interacts with double-stranded DNA (dsDNA) through an intercalation binding mode, exhibiting a binding constant (K) of 1.32 × 106M−1. Its amphiphilic nature and strong DNA affinity facilitate its localization within mitochondria in living cells and nuclei in apoptotic cells. Remarkably, within 30 min of COE-S3 staining, cell vitality can be discerned through real-time nuclear fluorescence imaging of apoptotic cells. COE-S3’s high DNA selectivity enables quantitative intracellular DNA analysis, providing insights into cell proliferation, differentiation, and growth. Our findings underscore COE-S3, with its strategically designed, shortened planar backbone, as a promising intercalative probe for DNA quantification and nuclear imaging.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Hui-Fang Cui,Tai-Bin Xu,Yu-Long Sun,An-Wei Zhou,Yu-Han Cui,Wei Liu,John H. T. Luong
Hairpin DNA (hpDNA) as a novel biobarcode was conjugated with gold nanoparticles (AuNPs) and a reporter DNA (rpDNA) to form hpDNA/AuNP/rpDNA nanoparticles for the detection of an oligonucleotide sequence associated with Helicobacter pylori as a model target. The rpDNA is complementary to about a half-portion of the target DNA sequence (tDNA). A capture DNA probe (cpDNA), complementary to the other half of the tDNA, was immobilized on the surface of a gold electrode. In the presence of tDNA, a sandwich structure of (hpDNA/AuNP/rpDNA)/tDNA/cpDNA was formed on the electrode surface. The differential pulse voltammetry (DPV) detection was based on [Ru(NH3)5(3-(2-phenanthren-9-yl-vinyl)-pyridine)]2+, an electroactive complex that binds to the sandwich structure by its intercalation with the hpDNA and the double-stranded DNA (dsDNA) of the sandwich structure. The several factors—high density of biobarcode hpDNA on the surface of AuNPs, multiple electroactive complex molecules intercalated with each hpDNA and dsDNA molecule, and the intercalation binding mode of the electroactive complex with the DNA sandwich structure—contribute to the DNA sensor with highly selective and sensitive sensing properties. The DNA sensor exhibited a detection limit of 1 × 10–15 M (i.e., 1 fM), the DNA levels in physiological samples, with linearity down to 2 × 10–15 M. It can differentiate even one single mismatched DNA from the complementary tDNA. This novel biobarcode-based DNA sensing approach should provide a general platform for development of direct, simple, repetitive, sensitive, and selective DNA sensors for various important applications in analytical, environmental, and clinical chemistry.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Ting Li,Shaojiao Song,Wei Lu,Yu Cao,Xin Zheng,Qiuyue Zhao,Hailong Wang
Owing to the high selectivity and binding affinity of coralyne (COR) with deoxyribonucleic acid (DNA)/ribonucleic acid, COR has been confirmed as an effective anticancer drug. However, a direct and sensitive detection assay for a DNA–COR–DNA structure with a portable instrument has not been reported because of the lack of a high-resolution sensor that can identify DNA–COR interaction at a single-molecule level. Here, specific Types I and II signals generated by a DNA–COR–DNA duplex were observed using a versatile α-hemolysin nanopore sensor. The Type I signal featuring a unique current pattern was preferentially selected via DNA sequence design and voltage control. Subsequently, the rapid and sensitive detection of COR was established with a detection limit of 0.1 nM. The pH environment, observed to influence the binding of the DNA–COR–DNA duplex, was significant for drug release controlling during targeted delivery process. Similar alkaloids were tested, and the nanopore sensor exhibited a high selectivity for COR. The results demonstrate a potential platform for investigating DNA-COR interaction and could be applied in the drug therapy and pharmacodynamic studies of COR.
✓ 已验证
产品关联
DOI
高影响因子
Biomaterials Science
·
2017
·Vol. 5
·期 7
·1279-1292
·
IF 4.21
Yawen Liu,Zhaozhu Zheng,He Gong,Meng Liu,Shaozhe Guo,Gang Li,Xiaoqin Wang,David L. Kaplan
The structure of DNA is susceptible to alterations at high temperature and on changing pH, irradiation and exposure to DNase. Options to protect and preserve DNA during storage are important for applications in genetic diagnosis, identity authentication, drug development and bioresearch. In the present study, the stability of total DNA purified from human dermal fibroblast cells, as well as that of plasmid DNA, was studied in silk protein materials. The DNA/silk mixtures were stabilized on filter paper (silk/DNA + filter) or filter paper pre-coated with silk and treated with methanol (silk/DNA + PT-filter) as a route to practical utility. After air-drying and water extraction, 50–70% of the DNA and silk could be retrieved and showed a single band on electrophoretic gels. 6% silk/DNA + PT-filter samples provided improved stability in comparison with 3% silk/DNA + filter samples and DNA + filter samples for DNA preservation, with ∼40% of the band intensity remaining at 37 °C after 40 days and ∼10% after exposure to UV light for 10 hours. Quantitative analysis using the PicoGreen assay confirmed the results. The use of Tris/borate/EDTA (TBE) buffer enhanced the preservation and/or extraction of the DNA. The DNA extracted after storage maintained integrity and function based on serving as a functional template for PCR amplification of the gene for zinc finger protein 750 (ZNF750) and for transgene expression of red fluorescence protein (dsRed) in HEK293 cells. The high molecular weight and high content of a crystalline beta-sheet structure formed on the coated surfaces likely accounted for the preservation effects observed for the silk/DNA + PT-filter samples. Although similar preservation effects were also obtained for lyophilized silk/DNA samples, the rapid and simple processing available with the silk–DNA–filter membrane system makes it appealing for future applications.
✓ 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）