SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Zhengfang Liang,Shuwei Xiao,Junhong Li,Guoli Zuo,Jihua Wei,Yuanxun Gong,Qianli Tang,Kai Zhang,Xianjiu Liao
Apurinic/apyrimidinic endonuclease 1 (APE1), a key enzyme in the base excision repair pathway, is frequently overexpressed in multiple cancers and has emerged as an important biomarker for early diagnosis and therapeutic response evaluation. In this work, a highly sensitive electrochemiluminescence (ECL) biosensor was developed for APE1 detection by integrating a PAM-regulated CRISPR-Cas12a activation mechanism with a multifunctional nanocomposite-modified electrode. A Ti₃C₂/AuNPs/CsPbBr₃ ternary nanocomposite was fabricated through ultrasonic assembly to provide enhanced conductivity, water stability, and probe-loading capacity. Meanwhile, a DNA duplex containing an AP site within the PAM region was engineered to regulate Cas12a activation through APE1-mediated cleavage and polymerase-assisted repair, thereby minimizing background interference and enabling highly specific signal generation. Ferrocene-labeled DNA tetrahedrons (TDNs) were further introduced as a spatially ordered probe scaffold to improve hybridization efficiency and facilitate Cas12a trans-cleavage–mediated signal release. This work presents three key innovations: (1) Construction of a Ti₃C₂/AuNPs/CsPbBr₃ nanocomposite that simultaneously enhances ECL stability, conductivity, and electron-transfer efficiency, addressing the inherent water-instability of perovskite emitters. (2) Coupling of a PAM-regulated APE1–KF repair mechanism with Cas12a activation to achieve highly specific AP-site recognition and selective signal generation. (3) Introduction of a TDN-based probe architecture that provides rigid spatial orientation and high-density probe loading, markedly improving hybridization efficiency and Cas12a trans-cleavage performance. Benefiting from these synergistic innovations, the biosensor achieves a detection limit of 6.28 × 10⁻⁸ U μL⁻¹ and a wide linear range (1 × 10⁻⁷ to 1 × 10⁻ 1 U μL⁻¹). The platform also demonstrates excellent specificity, reproducibility, and reliable performance in diluted serum, highlighting its promise for clinical and biochemical applications.
研究领域:
生物传感器
·
纳米技术
·
癌症诊断
·
电分析化学
·
分子生物学
·
CRISPR技术
✓ 已验证
产品关联
DOI
高影响因子
Nanoscale
·
2025
·
IF 5.1
Beibei Zhang,Xinyi Wang,Min Pan,Xinhao Li,Hong Wang,Yingwei Zhang
In this study, a novel dual-switch fluorescent biosensor based on a bipedal DNAzyme walker was developed for the highly sensitive detection of the tumour biomarker miRNA-141. By exploiting the typically elevated expression levels of miRNA-141 and apurinic/apyrimidinic endonuclease 1 (APE1) in tumour cells, an endogenous dual-gating system was constructed. The presence of both targets was required to initiate an enzyme-free strand displacement reaction, enabling target recycling and the subsequent release of bipedal DNAzyme walker components (D1/D2), which were efficiently isolated using streptavidin-modified magnetic beads. In the presence of Mg2+, the activated DNAzyme walker specifically cleaved fluorophore-labelled substrate strands on gold nanoparticles (AuNPs). Through repeated“walking–cleavage”cycles, fluorescence signals were progressively restored and amplified. By integrating AuNPs as carriers and magnetic separation technology, a cascade dual-amplification system based on non-enzymatic strand displacement and bipedal DNAzyme walking was established, markedly enhancing the detection sensitivity. The biosensor exhibited dual-switch-specific activation, effectively suppressing interference from non-target RNAs and other nucleases, and maintained a low background signal. It demonstrated excellent discrimination between tumour cells and normal cells, and showed good anti-interference performance in spiked serum samples. This study provides a new strategy for the development of high-precision tumour biomarker detection, offering potential for trace analysis in complex biological samples.
研究领域:
生物传感器
·
分子诊断
·
核酸化学
·
肿瘤标志物
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2024
·
IF 5.6
Changjiang Li,Yuqiang Hu,Tianzi Shi,Kejun Dong,Tongbo Wu
The expression level of human apurinic/apyrimidinic endonuclease 1 (APE1) is closely associated with the onset of various diseases, establishing it as a crucial clinical biomarker and a target in anti-cancer efforts. This study accomplished colorimetric and visual detection of APE1 by harnessing its endonuclease activity through catalytic hairpin self-assembly (CHA) and G-quadruplex/hemin DNAzyme. Optimization of the freedom degrees of the G-rich sequence significantly improved the detection performance of the strategy by influencing DNAzyme formation. Additionally, we replaced the signal reporting system with a molecular beacon to develop a fluorescence detection strategy, which served as an extension of the signal amplification system for validation and signal readout. The fluorescent probe method achieved a detection limit of 3.37 × 10 −4 U/mL, while the colorimetric method yielded a detection limit of 6.5 × 10 −3 U/mL, with a linear range spanning from 0.01 to 0.25 U/mL. Subsequently, the colorimetric approach effectively assessed APE1 activity in biological samples and facilitated the screening of APE1 activity inhibitors. Furthermore, this CHA/G-quadruplex/hemin DNAzyme strategy was adapted for the colorimetric detection of adenosine, showcasing its broad applicability across various biomarkers. The developed colorimetric analytical strategy represents a pivotal biosensing platform for diagnosing and treating diseases.
研究领域:
生物技术
·
分子生物学
·
分析化学
·
癌症研究
·
诊断学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ADVANCED FUNCTIONAL MATERIALS
·
2025
·
IF 19
Minghui Hu,Yingyu Zhang,Jie Zhang,Xianwei Zhang,Mengxin Zhang,Kangbo Liu,Lifeng Li,Zhidan Yu,Fei Zhang,Wancun Zhang
In vivo optical tumor molecular imaging encounters significant challenges in achieving adequate tumor specificity and sensitivity, largely attributed to off-tumor signal leakage and the relatively low expression levels of target molecules. Therefore, a double self-amplified programmable allosteric DNA nanomachine (named HPs-tFNA) is developed through two elaborately designed hairpin structures (HP1 and HP2) hybridized on tetrahedral framework DNA (tFNA), enabling rapid, specific, and sensitive tumor molecular imaging using the highly specific expression of apurinic/apyrimidinic endonuclease 1 (APE1) in the tumor cytoplasm as a stimulus-response target. In the presence of APE1, HP2 modifies two apurinic/apyrimidinic sites (AP sites), which can be specifically recognized and cleaved by APE1, releasing a significant number of cyclic sequences (cyclic-seq) and achieving initial APE1-assisted signal amplification. Subsequently, cyclic-seq hybridizes with HP1, inducing a conformational change that converts the stem-loop structure of HP1 to a linear form. This structural change facilitates the spatial separation of the fluorophore and quencher, thereby generating fluorescence signals. Furthermore, APE1 incises two AP sites within the HP1 loop region, resulting in the release of cyclic-seq. The released cyclic-seq can hybridize with additional HP1 to continuously amplify the fluorescence signal in a cyclic manner, thereby achieving the second round of signal amplification assisted by APE1. The experimental results of this study demonstrated that HPs-tFNA can achieve rapid in situ tumor molecular imaging and guide precise surgical excision in vivo, with superior spatial specificity. In particular, HPs-tFNA can effectively monitor drug resistance in neuroblastoma cells and stratify risk levels of neuroblastoma via plasma analysis.
研究领域:
生物医学工程
·
分子影像学
·
纳米医学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Guangxu Fang,Kai Wen,Qixuan Zhao,Yingxuan Chen,Yanbo Wang,Haobo Han,Quanshun Li
Postoperative tissue analysis is essential for the evaluation of surgical outcome in the tumor therapy. The identification of tumor biomarkers represents a supplementary approach to histochemical observation, facilitating the diagnosis of cancers at molecular level. Herein, a biosensor was developed by integrating an ATP-responsive aptazyme and its substrate on the surface of streptavidin magnetic beads to achieve the simultaneous detection of ATP and apurinic/apyrimidinic endonuclease 1 (APE1) within cancer cells. This strategy enabled the biosensor to respond to excessive ATP in cancer cells, thereby generating the primer. Meanwhile, the biosensor contained a double-stranded DNA with apurinic/apyrimidinic site, which allowed it to respond to APE1 and produce a padlock. The released primer and padlock triggered the rolling cycle amplification to yield an amplified fluorescent or colorimetric signal. The biosensor functioned as the “AND”-logic gate, effectively distinguishing cancer and non-cancerous cells. Moreover, the biosensor exhibited high specificity and sensitivity in discriminating the tumor and non-tumor tissues isolated from the patients bearing non-small cell lung cancer. Taken together, the biosensor showed the potential in clinical molecular diagnosis, serving as a promising supplementary approach for the postoperative tissue analysis.
研究领域:
生物医学工程
·
分子生物学
·
癌症诊断
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
Nano Today
·
2022
·
IF 18.962
Yecan Pan,Rui Weng,Linghao Zhang,Jing Qiu,Xinlu Wang,Guangqin Liao,Zhaohui Qin,Lingpu Zhang,Haihua Xiao,Yongzhong Qian,Xin Su
Cellular oxidative stress from effects of reactive oxygen species (ROS) and its induced molecular response play an important role in the pathogenesis of various diseases and important biological processes . Nanostructure-based fluorescent probes can simultaneously map the localization and abundance of ROS and its induced molecules in living cells. However, proximity-induced fluorophore interaction leads to low sensitivity and poor specificity. Here, we engineered intramolecular Orthogonal Reporters (iOR) based on DNA nanostructure scaffolds. Fluorescence reporters in iOR were precisely located with appropriate distance by simulation eliminating the undesired interaction of fluorophores. iOR functionalized with nuclear localization signal peptide is capable of imaging ROS and its related DNA repair enzyme (APE1) in the nucleus and cytoplasm with high sensitivity and specificity. iOR discloses the strong positive relation of ROS and APE1 as well as their synergistic regulation in living cells and tumor-bearing mice. This fact is first presented by direct fluorescence imaging. This work reported a new procedure for developing nanostructure-based molecular probes and would find broad applications in cellular biology and nanotechnology.
研究领域:
细胞生物学
·
纳米技术
·
分子生物学
·
生物医学工程
✓ 已验证
产品关联
DOI
高影响因子
MOLECULES
·
2023
·Vol. 28
·期 9
·3935
·
IF 4.6
Peng Lu,Xiangjian Cao,Jinghui Zheng,Ying Sun,Ziyu Tang,Meiping Zhao
As a major apurinic/apyrimidinic endonuclease and a redox signaling protein in human cells, APE1 plays a crucial role in cellular function and survival. The relationship between alterations of APE1 expression and subcellular localization and the initiation, development and treatment of various cancers has received extensive attention. However, comparing the in-vivo activity of APE1 in normal and cancerous breast live cells remains challenging due to the low efficiency of commonly used liposome transfection methods in delivering DNA substrate probes into human normal breast epithelial cells (MCF-10A). In this work, we develop a DNA/RNA hybrid-based small magnetic fluorescent nanoprobe (25 ± 3 nm) that can be taken up by various live cells under magnetic transfection. The D0/R-nanoprobe demonstrates an outstanding specificity toward APE1 and strong resistance to the cellular background interference. Using this nanoprobe, we are not only able to visualize the intracellular activity of APE1 in breast ductal carcinoma (MCF-7) live cells, but also demonstrate the APE1 activity in MCF-10A live cells for the first time. The method is then extended to observe the changes in APE1 levels in highly metabolically active neuroendocrine cells under normal conditions and severe attacks by reactive oxygen species in real-time. The fluorescent nanoprobe provides a useful tool for studying the dynamic changes of intracellular APE1 in normal or cancerous live cells. It also displays the potential for visible and controllable release of miRNA drugs within live cells for therapeutic purposes.Keywords:apurinic/apyrimidinic endonuclease 1 (APE1);fluorescent nanoprobe;live cell fluorescence imaging;human normal breast epithelial cells (MCF-10A);human breast adenocarcinoma cells (MCF-7)
研究领域:
癌症生物学
·
细胞生物学
·
纳米技术
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2026
·
IF 7.3
Fei Zhang,Yingyu Zhang,Xianwei Zhang,Qionglin Wang,Na Zhao,Jianying Ren,Man Zhang,Peng Wang,Mengxin Zhang,Wancun Zhang
Abstract Mitochondrial apurinic/apyrimidinic endonuclease 1 (APE1) mislocalization is closely linked to tumor progression and prognosis. Yet, its ubiquitous distribution across mitochondria, cytoplasm, and nucleus makes specific in situ imaging of mitochondrial APE1 highly challenging. Herein, we fabricate a sequentially activated biosensor (C-12S-tFNA) with 12S rRNA-gated spatial regulation of APE1 cleavage site (apurinic/apyrimidinic site, AP site) accessibility, where cytochrome c (cyt c) acts as the targeting motif to drive precise mitochondrial enrichment. In the initial state, the AP site is structurally locked to prevent nonspecific cleavage by extramitochondrial APE1. After entering the mitochondria, mitochondrial 12S rRNA, which is consistently expressed across diverse cell types, hybridizes with the AP-site-containing strand via base complementarity, leading to AP site unlocking. The exposed AP site is subsequently cleaved by mitochondrial APE1, separating the fluorophore-quencher pair and generating a fluorescent signal. Benefiting from the 12S rRNA-gated design, false-positive signals can be effectively suppressed. Experimental findings demonstrate that C-12S-tFNA exhibits high sensitivity (LOD: 0.002 U/mL) and exceptional specificity for mitochondrial APE1. Notably, it distinguishes high-risk from low-risk neuroblastoma (NB) in vivo without the need for surgically resected or biopsied tissue, while exhibiting favorable biocompatibility. Collectively, C-12S-tFNA enables noninvasive, activity-based mitochondrial APE1 imaging and holds promise for improving precision risk stratification in NB.
✓ 已验证
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Yunlong Wang,Jie Zhou,Yan Dong,Peng Wang,Ruiyang Zi,Honglan Zhang,Yingqiang Luo,Yan Zhang,Teng Wang,Yuhan Chen,Biao Hu,Houjie Liang,Zhihao Wei,Xiang Zhao
Constructing precise delivery systems to enhance the antitumor efficacy of drugs represents a pressing clinical practice need. The characteristics of suitable physiological stability, flexible tunable size, and highly programmable architecture make functional DNA scaffolds an ideal solution to this requirement. Here, we designed DNA oligonucleotide probes Y 1 , Y 2 and Y 3 to construct a smart-responsive Y-shaped DNA scaffold drug conjugate (sYDD). This sYDD incorporates two AS1411 aptamers, an apurinic/apyrimidinic (AP) site, a B-cell lymphoma-2 (Bcl-2) antisense oligonucleotide (ASO) and a doxorubicin (DOX)-carrying sequence. The Bcl-2 ASO and DOX within sYDD could be responsively released in tumor cells that exhibited high expression of apurinic/apyrimidinic endonuclease 1 (APE1) and microRNA-21 (miR-21). And the in vitro and in vivo experiments demonstrated that sYDD exhibited substantial antitumor efficiency. Moreover, the flow cytometry and RNA sequencing results proved that the proposed sYDD effectively activated immune responses. This strategy provides an alternative approach for designing synergistic chemo-gene drug conjugates, which facilitates the development of precision tumor treatment modalities.
研究领域:
药学
·
生物技术
·
癌症研究
·
分子生物学
·
纳米医学
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Yanting Feng,Zewei Luo,Juan Wu,Zhijun Huang,Lu He,Yaqiang Feng,Xian Li,Xudong Zhang,Yonghui Tian,Yongxin Li,Yixiang Duan
Present DNA logic platforms mainly focused on distinguishing a specific kind of cancer cell from several other types of cell lines, while limited efforts have been paid to high-throughput recognition of cancer cells. A new DNA logic platform coupled with multivariate statistical analysis was developed to classify seven kinds of cancer cells. Four confined DNA circuits containing localized catalytic hairpin assembly (LCHA) and apurinic/apyrimidinic endonuclease 1 (APE1)-assisted circuits were conjugated to a protein of streptavidin (SA) for DNA logic platform. The mismatch minimizes the circuit leakage of the confined DNA circuits by inhibiting the incorrectly intramolecular structure switch. Benefitting from the spatial-confined effect, the DNA logic platform as a modified AND logic gate sensitively responded to the dual intracellular biomarkers, thereby generating strong dual fluorescent output signals in living cells. Based on the fluorescent cell imaging, multidimensional data collected from 50 individual cells were concatenated as a dataset for multivariate statistical analysis. Principal component analysis (PCA) shows distinct clustering among the seven kinds of cancer cell lines. A random forest algorithm obtained the specificity and sensitivity of 100%. Therefore, the DNA logic platform coupling with multivariate statistical analysis shows promising potential for cancer diagnosis and treatment.
研究领域:
生物技术
·
癌症生物学
·
分子生物学
·
生物信息学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）