SENSORS AND ACTUATORS B-CHEMICAL
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2020
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IF 7.1
Yuqiang Hu,Zhen Zhang,Weicong Ye,Wei Zhang,Minghao Hu,Wenqian Yuan,Hongbo Wang,Xianjin Xiao,Tongbo Wu
Human apurinic/apyrimidinic endonuclease 1 (APE1) plays a crucial role in DNA repair and gene regulation. The abnormal variations of the concentration of APE1 in the human blood and tissue cells are highly correlated to various diseases. Thus, APE1 can be used as a biomarker to aid clinical diagnosis of diseases. Detection of APE1 activity with high sensitivity and selectivity simultaneously is difficult to achieve. Here we provide a DNA structure-mediated fluorescent biosensor to solve the above problem. Upon the existence of APE1, the apurinic/apyrimidinic site will be cleaved, and the terminal structure of the hairpin DNA substrate will change. Then the terminal deoxynucleotidyl transferase and endonuclease IV could provide dual fluorescent signal amplification. Through the ingenious structure design of the biosensor, we have improved the reactivity of APE1 and minimized the interference of other enzymes. The results indicate that the detection limit for APE1 is as low as 1.7 × 10 −6 U/mL. The biosensor has been successfully applied to the detection of APE1 in real biological samples and the screening of APE1 inhibitors.
研究领域:
生物化学
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分子生物学
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医学诊断
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DOI
高影响因子
MICROCHEMICAL JOURNAL
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2025
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IF 5.1
Jihua Wei,Pengyuan He,Xudong Xin,Yuanxun Gong,Jiayi Zhang,Longjian Huang,Kai Zhang,Xianjiu Liao
A label-free, isothermal, surface-confined electrochemiluminescence assay for apurinic/apyrimidinic endonuclease-1 (APE1) is presented, in which target-triggered isothermal amplification is coupled to a bipedal DNA walker on a PEI-Ru(dcbpy)₃ 2+ /AuNPs@Ti₃C₂ (MXene) transducer to provide high signal with low background. Under optimized conditions, a log-linear range of 1.0 × 10 −3 –1.0 × 10 5 U·L −1 and a limit of detection of 5.62 × 10 −3 U·L −1 were obtained. Applicability in complex media was demonstrated in diluted human serum, yielding recoveries of 96.56–104.51 % with RSD <5 %. Selectivity controls against common nucleases and proteins showed minimal interference, and stepwise electrochemical characterization supported successful interface construction. Feasibility of the nucleic-acid modules and pathway was verified by native and denaturing PAGE, and material identity/localization was corroborated by updated EDS mapping with enhanced Ru/C clarity. By integrating dual amplification on a conductive MXene/Ru interface in a compact, label-free format, this platform enables activity-unit quantitation of APE1 in biologically relevant samples and can be adapted to other DNA-repair enzymes by reprogramming recognition sequences.
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高影响因子
ANALYTICAL CHEMISTRY
·
2023
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IF 7.4
Junqiu Zhai,Fanglin Huang,Yunyun Yang,Xiaoxin Liu,Tiangang Luan,Jiewei Deng
Pollutant exposure causes a series of DNA damage in cells, resulting in the initiation and progression of diseases and even cancers. An investigation of the DNA damage induced by pollutants in living cells is significant to evaluate the cytotoxicity, genotoxicity, and carcinogenicity of environmental exposure, providing critical insight in the exploration of the etiologies of diseases. In this study, we develop a repair enzyme fluorescent probe to reveal the DNA damage caused by an environmental pollutant in living cells by single-cell fluorescent imaging of the most common base damage repair enzyme named human apurinic/apyrimidinic endonuclease 1 (APE1). The repair enzyme fluorescent probe is fabricated by conjugation of an APE1 high affinity DNA substrate on a ZnO2 nanoparticle surface to form a ZnO2@DNA nanoprobe. The ZnO2 nanoparticle serves as both a probe carrier and a cofactor supplier, releasing Zn2+ to activate APE1 generated by pollutant exposure. The AP-site in the DNA substrate of the fluorescent probe is cleaved by the activated APE1, releasing fluorophore and generating fluorescent signals to indicate the position and degree of APE1-related DNA base damage in living cells. Subsequently, the developed ZnO2@DNA fluorescent probe is applied to investigate the APE1-related DNA base damage induced by benzo[a]pyrene (BaP) in living human hepatocytes. Significant DNA base damage by BaP exposure is revealed, with a positive correlation of the damage degree with exposure time in 2–24 h and the concentration in 5–150 μM, respectively. The experimental results demonstrate that BaP has a significant effect on the AP-site damage, and the degree of DNA base damage is time-dependent and concentration-dependent.
研究领域:
环境生物学
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分子生物学
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毒理学
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DOI
PubMed
高影响因子
TALANTA
·
2026
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IF 6.7
Junfeng Peng,Jizhan Mai,Wanting Shi,Yun Liu,Lang Zhang,Chunrong Li,Jiahong Lei
Developed the APE1-assisted signal amplification strategy for ultrasensitive Hg 2+ detection. • Detection limit as low as 2.7 pM, far below the EPA regulatory level (10 nM). • Homogeneous one-step assay completed within 40 minutes with simple operation. • Excellent selectivity against competing metal ions and reliable recoveries (95.0–104.8%). Mercury (Hg 2+ ) is a highly toxic heavy metal pollutant whose environmental persistence and biomagnification through the food chain pose serious risks to ecosystems and human health. Hence, there is considerable interest in developing sensitive, selective, and convenient methods for its detection. Herein, we report a novel fluorescent sensor for ultrasensitive and rapid analysis of Hg 2+ , based on apurinic/apyrimidinic endonuclease 1 (APE1)-assisted signal amplification. The sensor comprises a hairpin reporter probe (RP) containing an abasic (AP) site and dually labeled with CY3 and BHQ2, along with a T-rich helper probe (Helper). Upon addition of Hg 2+ , the Helper hybridizes to the T-rich loop of RP through T-Hg 2+ -T coordination, forming a duplex that enables APE1 to cleave the AP site efficiently. This cleavage releases a short CY3-labeled fragment, generating a strong fluorescence signal. Simultaneously, both Hg 2+ and the Helper are regenerated, allowing successive cleavage cycles to occur and thereby achieving multiple signal amplification. The entire assay is performed in a homogeneous solution in a single step, with a total reaction time of 40 min. Under optimal conditions, the sensor achieves a detection limit as low as 2.7 pM and a linear dynamic range of 0.01–10.0 nM, and it exhibits excellent selectivity against common interfering metal ions. Spike-recovery tests in real water samples gave recoveries between 95.0% and 104.8%, showing no significant deviation from results obtained by ICP-MS. This strategy is operationally simple, cost-effective, and highly sensitive, offering a promising tool for rapid screening of trace Hg 2+ in environmental and food safety monitoring. Download: Download high-res image (151KB) Download: Download full-size image
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2023
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IF 8.4
Mengxu Sun,Yanxin Zhang,Juan Xie,Ya Zhang,Ting Huang,Minmin Li,Jin-Xiang Chen,Zong Dai,Jun Chen
Developing simple, rapid and sensitive strategies for miRNA analysis is extremely important for diseases early diagnosis. Herein, an APE1-mediated isothermal target cycling amplification system combined with magnetic separation was established for label-free and rapid detection of miRNA. As a proof-of-concept, miR-1246 was selected as research model. In the detection system, when the miR-1246 is present, it hybridizes with AP probes of AP-MBs to form a double-stranded, in which the APE1 recognizes specific AP site of double-stranded and cleaves the AP probes, releasing a sequence containing a G-quadruplex (G4). After the cleavage, target miR-1246 can be released to hybridize with another AP of AP-MBs, triggering a continues cleavage reaction. After magnetic separation, label-free analysis of miRNA was achieved by measuring the fluorescence of G4/ThT. Due to the high efficiency and specificity of APE1 toward AP sites in dsDNAs, this method exhibits high specificity and sensitivity with the capability of distinguishing miRNAs with single-base mismatch. The system could detect miRNA with high sensitivity within 2 h, and the limit of detection (LOD) was calculated as 25.6 fM. Furthermore, high accuracy has been achieved through recovery experiments and successful attempts has been made in applying the approach to detect miR-1246 in serum samples from breast cancer patients and normal people. This method is expected to be an effective tool for miRNA-related research and clinical early diagnosis.
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高影响因子
ANALYTICAL CHEMISTRY
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2025
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IF 6.7
Lanlan Chen,Jingjing Lai,Siqi Dong,Wenjun Liu,Ximei Zhang,Huanghao Yang
Evaluating tumor radiosensitivity is beneficial for the prediction of treatment efficacy, customization of treatment plans, and minimization of side effects. Tracking the mitochondrial DNA (mtDNA) repair process helps to assess tumor radiosensitivity as mtDNA repair determines the fate of the cell under radiation-induced mtDNA damage. However, current probes developed to monitor levels of DNA repair enzymes suffered from complex synthesis, uncontrollable preparation, limited tumor selectivity, and poor organelle-targeting ability. Especially, the correlation between mtDNA repair activity and inherent radiosensitivity of tumors has not yet been explored. Here, we present a mitochondria-targeted DNA-based nanoprobe (TPP-Apt-tFNA) for in situ monitoring of the activity of the mtDNA repair enzyme and evaluating tumor radiosensitivity. TPP-Apt-tFNA consists of a DNA tetrahedral framework precisely modified with three functional modules on each of the three vertexes, that is, the tumor cell-targeting aptamer, the mitochondrion-targeting moiety, and the apurinic/apyrimidinic endonuclease 1 (APE1)-responsive molecule beacon. Once selectively internalized by tumor cells, the nanoprobe targeted the mitochondrion and specifically recognized APE1 to activate fluorescence, allowing the observation of mtDNA repair activity. The nanoprobe showed elevated APE1 levels in the mitochondria of tumor cells under oxidative stress. Moreover, the nanoprobe enabled the illumination of different levels of APE1-mediated mtDNA repair activity in different cell cycle phases. Furthermore, using the nanoprobe in vitro and in vivo, we found that tumor cells with high activity of mtDNA repair, which allowed them to recover from radiation-induced mtDNA lesions, had low sensitivity to radiation and an unsatisfactory radiotherapy outcome. Our work provides a new imaging tool for exploring the roles of mtDNA repair activity in diverse biological processes and for guiding tumor radiation treatment.
研究领域:
生物化学
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分子生物学
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医学影像学
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肿瘤学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2026
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IF 4.1
Yang Chan,Yan Zihan,Qin Yanru,Wei Ying,He Ziyi,Mo Liuting,Lin Weiying
Apurinic/apyrimidinic endonuclease 1 (APE1), a critical enzyme in the base excision repair pathway, is a potential biomarker for numerous cancer types. Traditional APE1 activity detection methods are hindered by complex protocols, expensive labeling requirements, and vulnerability to environmental factors. Herein, we develop a label-free ratiometric fluorescent biosensor (TCHG) that integrates APE1-mediated catalytic hairpin assembly (CHA) amplification with split G-quadruplex/malachite green (MG)–mediated aggregation-induced emission (AIE) signaling for sensitive activity analysis of APE1. Our system operates through APE1’s specific cleavage of apurinic/apyrimidinic (AP) site-containing double-stranded substrates, which releases trigger strands that initiate a CHA cascade. This cascade drives the assembly of split G-quadruplex structures. Upon reconstruction, the complete G-quadruplex binds MG, generating amplified near-infrared fluorescence with AIE characteristics. Simultaneously, Hoechst 33258 (HOE) embedded within double-stranded DNA provides a stable reference signal, enabling self-calibrated ratiometric output that effectively minimizes signal fluctuations from instrumental and environmental variations. The TCHG biosensor demonstrates high selectivity for APE1 with a detection limit of 2.4 × 10 –3 U/mL. Importantly, it maintains reliable analytical performance in complex biological matrices, including fetal bovine serum and various cell lysates, successfully differentiating endogenous APE1 activity between normal and cancer cells. The TCHG system offers a tool for APE1 activity analysis in biological samples, with potential applications in biomedical research. Graphical abstract
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高影响因子
CHEMICAL ENGINEERING JOURNAL
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2024
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IF 13.3
Mengxin Zhang,Yingyu Zhang,Xianwei Zhang,Qionglin Wang,Kangbo Liu,Muchun Yu,Lifeng Li,Zhidan Yu,Huan Zhao,Weyland Cheng,Wancun Zhang
Specific imaging of tumor cells with high sensitivity is urgently needed for evaluating the risk of tumor metastasis and progression. However, traditional tumor cell imaging strategies are still restricted by limited tumor specificity and sensitivity due to off-tumor signal leakage and a lower content of the targets. Therefore, a self-powered and self-recycled strategy based on molecular beacon (MB) hybridized on tetrahedral framework DNA (E-MB-tFNA) using the endogenous enzyme, apurinic/apyrimidinic endonuclease 1 (APE1), was rationally developed for tumor cell-specific molecular imaging with superior spatial specificity and improved sensitivity. The apurinic/apyrimidinic site (AP site) can be specifically recognized and cleaved by APE1, and was modified on the stem and loop region of E-MB-tFNA. The resultant E-MB-tFNA can be specifically recognized and cleaved by APE1 in the cytoplasm, which is overexpressed in tumor cells compared to normal cells. In particular, once APE1 cleaves the AP site on the stem region (double-stranded DNA) of E-MB-tFNA, the self-powered and self-recycled sequences modified on the stem region of E-MB-tFNA is freed to hybridize with the loop region of E-MB-tFNA, prompting APE1 to further hydrolyze the E-MB-tFNA loop region and generate fluorescence signals. In addition, the self-powered and self-recycled sequences can be hybridized with other E-MB-tFNA to continuously and cyclically amplify the fluorescence signal. The experimental results of this study showed that the E-MB-tFNA has the capability to monitor drug resistance and achieve guided precision surgical excision in vivo with superior spatial specificity and improved sensitivity. In particular, E-MB-tFNA can realize the risk classification of neuroblastoma patients in clinical plasma.
研究领域:
生物医学工程
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分子生物学
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肿瘤学
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高影响因子
ACS Applied Materials & Interfaces
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2023
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IF 9.5
Zening Huang,Kaixiang Xu,Lijuan Zhao,Li-e Zheng,Nana Xu,Caixia Yan,Xingjiang Hu,Qiao Zhang,Jian Liu,Qingwei Zhao,Yaokun Xia
The development of a strategy for imaging of glutathione (GSH) and apurinic/apyrimidinic endonuclease 1 (APE1) in an organism remains challenging despite their significance in elaborating the correlated pathophysiological processes. Therefore, in this study, we propose a DNA-based AND-gated nanosensor for fluorescence imaging of the GSH as well as APE1 in living cells, animals, and organoids. The DNA probe is composed of a G-strand and A-strand. The disulfide bond in the G-strand is cleaved through a GSH redox reaction, and the hybridization stability between the G-strand and A-strand is decreased, leading to a conformational change of the A-strand. In the presence of APE1, the apurinic/apyrimidinic (AP) site in the A-strand is digested, producing a fluorescence signal for the correlated imaging of GSH and APE1. This nanosensor enables monitoring of the expression level change of GSH and APE1 in cells. Additionally, we illustrate the capability of this “dual-keys-and-locked” conceptual methodology in achieving specific tumor imaging when GSH and APE1 are present simultaneously (overexpressed GSH and APE1 in tumor cells) with improving tumor-to-normal tissue ratio in vivo. Furthermore, using this nanosensor, the GSH and APE1 also are visualized in organoids that recapitulate the phenotypic and functional traits of the original biological specimens. Overall, this study demonstrates the potential of our proposed biosensing technology in investigating the roles of various biological molecules involved in specific diseases.
研究领域:
生物医学工程
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分子生物学
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细胞生物学
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癌症研究
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纳米技术
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2026
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IF 11.8
Ting Huang,Ziyue Qiu,Siyuan Tan,Luyin Zhang,Jing Peng,Simin Liu,Xiaoming Lyu,Zong Dai,Jun Chen,Jin-Xiang Chen
Precise imaging of intracellular biomarkers is crucial for accurate disease diagnosis and mechanistic insight. Although confined catalytic hairpin assembly (CCHA) can accelerate reaction kinetics, its performance is frequently compromised by nonspecific amplification, resulting in high background signals and limited imaging contrast. Here, we develop a tumor-upregulated endogenous apurinic/apyrimidinic endonuclease 1 (APE1)-triggered CCHA (AP-CCHA) system based on a DNA nanowire for selective microRNA (miRNA) imaging in living tumor cells. Using miR-21 as a proof-of-concept model, the mechanistic analyses demonstrated that conventional CCHA suffers from significant nonspecific signal amplification during both in vitro assembly and storage. For the proposed AP-CCHA system, we incorporated an APE1-activatable AP-H1 probe that improves molecular stability, suppresses nonspecific amplification, and consequently enhances the signal-to-noise ratio while preserving high reaction kinetics. As a result, the AP-CCHA system delivers a 3.5-fold improvement in sensitivity compared with CCHA and achieves an 18.2 pM detection limit for miR-21. Leveraging the elevated cytoplasmic APE1 levels in tumor cells, AP-CCHA remains quiescent in normal cells but is selectively activated in cancer cells, enabling high-contrast miRNA discrimination and precise in vivo tumor imaging, and effectively minimizing false positives. By integrating confinement-enhanced kinetics with endogenous enzyme activation, this strategy enables rapid, highly sensitive, and high-contrast biomarker imaging. The AP-CCHA platform thus provides a versatile analytical tool for nucleic acid-based diagnostics with strong potential for precision therapeutics and drug development applications.
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
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2026
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IF 7.3
Junli Liu,Zhijuan Duan,Xianyan Gong,Fan Xia,Fujian Huang
Abstract Apurinic/apyrimidinic endonuclease 1 (APE1) is a clinical biomarker whose small molecular size hinders direct detection by conventional nanopores (30 nm). Here, we report a nanopipette platform integrating APE1-driven enzymatic recycling amplification with DNA tetrahedron−streptavidin (TDN−SA) complex-based resistive-pulse reporting to overcome this size limitation. APE1 specifically cleaves hairpin DNA probes, generating output strands that displace biotin-modified DNA oligo from the TDN−SA complex, reducing translocation event frequency in 30 nm nanopipettes. This signal modulation enables APE1 quantification with a detection limit of 10−5 U/mL and a linear range spanning 7 orders of magnitude (10−5 to 10 U/mL). The method discriminates APE1 activity between normal and cancer cell lines (HeLa, MCF-7, A549 vs. 293T, MCF-10A), tracks oxidative stress-induced elevation (0−4 h H2O2 treatment), and reveals cell cycle-dependent regulation (lowest in G0/G1, peaking in S phase). Furthermore, the system evaluates APE1 inhibition by 7-nitroindole-2-carboxylic acid in inflammatory macrophages, demonstrating its utility for drug efficacy assessment. This label-free platform overcomes the protein size constraint of traditional nanopore methods, offering a versatile tool for profiling low-abundance protein biomarkers in native cellular contexts.
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高影响因子
CHEMICAL ENGINEERING JOURNAL
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2025
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IF 13.2
Wancun Zhang,Zhenzhen Tian,Pin Zhang,Lifeng Li,Zhidan Yu,Meng Sun,Kangbo Liu,Ligong Hou,Jie Zhang,Mengxin Zhang,Xianwei Zhang
Clinical practice has demonstrated that the absence of in vivo drug resistance monitoring strategies for neuroblastoma (NB) contributes to relapse rates exceeding 60 % among children with high-risk NB. These patients frequently miss the optimal therapeutic window for treatment, leading to poor prognoses and ultimately fatal outcomes. Herein, a novel strategy (APII-tFNA) was developed by targeting apurinic/apyrimidinic endonuclease 1 (APE1) and miR-514a for rapidly and spatially in vivo imaging of NB and monitoring NB drug resistance. Both APE1 and miR-514a have been rigorously validated as critical NB drug resistance markers at the clinical NB tissue level. APII-tFNA, the APE1 cyclically cleaved and miR-514a self-recycled programmable allosteric DNA nanomachine, operates via an AND-gated manner. APII-tFNA can hybridize with miR-514a, leading to the formation of the 514a-APII-tFNA complex. Subsequently, the apurinic/apyrimidinic site (AP site) within the 514a-APII-tFNA complex can be specifically cleaved by APE1, leading to spatial separation between fluorophore and quencher, and configurational changes in 514a-APII-tFNA. Simultaneously, APE1 and miR-514a can enter the next reaction cycle to generate more fluorescence signals, thereby enabling the highly sensitive detection of targets. Experimental results demonstrated that this responsive nanomaterial achieved a limit of detection (LOD, 3σ) of 0.25 pM for in vivo molecular imaging of NB drug resistance marker and was able to guide precise surgical excision with superior spatial specificity. This study presents a nanomaterial platform for in vivo molecular imaging of NB and monitoring NB drug resistance, with great potential for improving the survival rate of children with NB.
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高影响因子
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