ACS Nano
·
2025
·
IF 16
Xiaohui Zhan,Yujia Jiang,Zhiyu Li,Xiaoqin Hu,Fang Lan,Binwu Ying,Yao Wu
The screening and monitoring of microRNAs as cancer molecular biomarkers is clinically significant, but traditional methods lack sufficient sensitivity, accuracy, and convenience. The CRISPR-colorimetric lateral flow assay (CLFA) integration offers a promising and efficient solution; however, cumbersome preamplification and poor quantification hinder clinical adoption. In this study, we developed a one-step isothermal CRISPR-Cas cascaded sensing system that is preamplification-free. At its core is a designed and selected split DNA tetrahedron activator, employing spatiotemporal-hierarchy mechanisms to precisely bidirectionally drive the kinetics of two Cas enzymes, accelerating the activation of Cas13a while delaying the initiation of Cas12a, to achieve optimal balance. This system enables ultrasensitive, single-step, single-tube, and rapid detection of a cervical cancer relative biomarker, microRNA-21, achieving a limit of detection of 38 aM with a broad linear range. The CRISPR system is further integrated with CLFA enhanced by ultrathin platinum-protected gold nanolabels (Au@Pt, also named Au@s-Pt), along with a smartphone equipped with dual convolutional neural network models (YOLO v5 and MobileNet v3), enabling more precise, rapid quantification of target miRNA. Using this integrated platform, miRNA-21 levels in cervical cancer and precancerous samples can be accurately quantified with approximately 30 min at low cost and without the need for large, sophisticated instruments, with results showing good concordance with quantitative real-time polymerase chain reaction. This platform provides an efficient, highly sensitive, user-friendly, and quantifiable point-of-care testing solution.
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PubMed
高影响因子
Materials Today Bio
·
2026
·
IF 10.2
Huanqing Zhu,Haonan Yu,Mengxin Huang,Panjie Sun,Yuanfang Tang,Zhanyi Zhang,Puming He,Youying Tu,Bo Li
Atopic dermatitis (AD) is an inflammatory skin disease characterized by a complex pathogenesis. Current clinical treatments exhibit several limitations, including low efficacy and considerable side effects, prompting a growing interest in novel therapeutic strategies. In this study, we developed a microneedle-assisted dual-nano drug delivery strategy that targets both oxidative stress and Janus kinase 1 (JAK1) expression, aiming for enhanced therapy of AD. Self-assembling nanoparticles (NPs) incorporating epigallocatechin gallate (EGCG) and lactoferrin were developed to effectively modulate oxidative stress. Concurrently, CRISPR-CasRx technology was employed to silence JAK1, and nanoparticles were constructed by encapsulating CasRx and siRNA within poly (β-amino ester) (PBAE). Both types of NPs were successfully delivered to skin lesions in AD mice through microneedles, either individually or combined, resulting in significant alleviation of symptoms. Treatment reduced dermatitis severity scores and splenomegaly, decreased epidermal thickness and mast cell infiltration, and increased collagen fiber content. Additionally, levels of inflammatory cytokines (IL-1β, IL-4, IL-13) and oxidative DNA damage marker (8-OHdG) in dorsal skin tissues were lowered, accompanied by decreased serum levels of thymic stromal lymphopoietin (TSLP) and IgE. The therapy also suppressed JAK1 expression and activated the Nuclear factor erythroid 2-related factor 2 (Nrf2)/Heme oxygenase 1 (HO-1) antioxidant pathway, except for PBAE-plasmid NP-loaded microneedles, which did not significantly increase HO-1 expression. Notably, microneedles containing both NPs exhibited superior efficacy in reducing IL-1β and JAK1 levels and upregulating Nrf2 expression compared to formulations with only one NP type. This innovative combination strategy demonstrates promise as a novel therapeutic approach for AD.
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PubMed
高影响因子
ACS Sensors
·
2026
·
IF 10.9
Tianchi Zhuang,Yan Long,Yongtong Xu,Qiumei Li,Zhiming Liu,Xueshan Zhong,Dawei Wu,Yu Ma,Wenzhe Su,Xiaokang Li,Zunfu Ke,Cheng Guo
Early and accurate diagnosis of Chikungunya virus (CHIKV) infection is critical for controlling its outbreaks. CRISPR/Cas-based detection offers promise for pathogen identification, yet one-step CRISPR/Cas systems are limited by suboptimal sensitivity, field-deployability, and adaptability to complex clinical samples, hindering their use in rapid outbreak response. Here, we developed a CRISPR/Cas13a-based One-Step System for Rapid Detection of Emerging Viruses (CRISPR-CORE) and applied it during the CHIKV outbreak in Guangdong Province, China. Multidimensional optimizations enabled the CRISPR-CORE system to achieve a limit of detection of 5 copies/μL within 40 min. An extraction-free RNA release protocol for CHIKV in blood samples and a premixed reagent approach were implemented. Furthermore, a portable fluorescence detector was used to enhance user-friendliness in point-of-care (POC) settings. The clinical CHIKV genomic information was identified through hybrid capture sequencing, informing the design of highly specific CRISPR RNA (crRNA). Clinical validation across three regions yielded 92.6% sensitivity and 100% specificity, underscoring the applicability and reliability of the CRISPR-CORE system. Our system demonstrates its suitability for CHIKV outbreak detection. It facilitates rapid and POC testing for emerging viruses in resource-limited settings. Furthermore, it provides a universal strategy for the prevention and control of infectious diseases.
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高影响因子
Analytical Methods
·
2025
·
IF 2.6
Zixun Wang,Dong Zhang,Yuxuan Wu,Xu Ye,Yayun Qian,Fengsong Chen
The low early diagnosis rate of hepatocellular carcinoma (HCC) severely impacts patient prognosis, making the development of highly sensitive and specific early diagnostic technologies crucial. MicroRNA-221 (miR-221), an aberrantly overexpressed biomarker in HCC, holds significant diagnostic potential. This paper constructed a capillary surface-enhanced Raman scattering (SERS) sensing platform utilizing CRISPR/Cas13a trans-cleavage and double-shell gold-silver nanorods (DS Au-AgNRs) to detect serum miR-221 in HCC patients. DS Au-AgNRs were synthesized and assembled onto aminated capillaries, followed by conjugation of Cy5-labeled single-stranded DNA (ssDNA) to the DS Au-AgNR surface via Au–S bonds. In the presence of miR-221, activated CRISPR/Cas13a trans-cleavage cleaves the ssDNA, releasing Cy5 from the sensor surface and diminishing the SERS signal, enabling miR-221 quantification. The synthesized DS Au-AgNRs exhibit uniform morphology and size, are uniformly distributed on the capillary, and form numerous “hotspots”, thereby significantly enhancing the SERS signal. According to the characteristic peak of Cy5 at 1074 cm−1, a linear relationship is established between the log concentration of miR-221 and the measured SERS intensity (y = −3527.97 × −35369.60, R2 = 0.97767), with a LOD as low as 4.17 × 10−17 M. The sensor demonstrated high specificity and high sensitivity, and its capacity to detect miR-221 expression aligned with qRT-PCR results when analyzing serum samples, confirming that hepatocellular carcinoma patients exhibited significantly higher miR-221 levels compared to healthy individuals. The capillary SERS sensor thus provides an accurate and convenient approach for early HCC diagnosis.
研究领域:
生物医学工程
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分子诊断
·
纳米生物技术
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癌症生物标志物
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表面增强拉曼散射（SERS）
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CRISPR技术
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PubMed
高影响因子
Sensors and Actuators Reports
·
2025
·
IF 7.6
Yanli Sun,Hongying Li,Tao Wang,Kai Zhang,Xueliang Wang
MicroRNA-17 (miR-17) is a critical biomarker linked to various cancers and diseases, underscoring the need for sensitive and selective detection methods. Herein, an ultra-sensitive electrochemiluminescent (ECL) sensor for miR-17 detection utilizing MOF-derived porous CuFe 2 O 4 nanospheres and CRISPR/Cas13a trans-cleavage-triggered hybridization chain reaction (HCR) amplification was presented. The porous CuFe 2 O 4 nanospheres possessed a high surface area and excellent catalytic activity, significantly enhancing ECL emission. To the best of our knowledge, this is the first report demonstrating that MOF-derived CuFe 2 O 4 can effectively enhance the electrochemiluminescence of Ru(bpy) 3 2 ⁺ for miR-17 detection. The CRISPR/Cas13a system provided miR-17 recognition and trans-cleavage activity, triggering HCR for signal amplification. Under optimized conditions, the biosensor exhibited a wide linear detection range from 20 aM to 5000 aM, with a detection limit of 9.23 aM. The system discriminated miR-17 from closely related miRNAs with negligible cross-reactivity, which demonstrated its outstanding selectivity. Practical applicability was confirmed by successful quantification of miR-17 in diluted human serum which achieved recovery rates between 95.2% and 104.8%. The results validated the biosensor’s potential for early disease diagnosis and clinical monitoring of miR-17-associated disorders.
研究领域:
生物传感器
·
分析化学
·
纳米材料
·
分子诊断
·
癌症研究
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高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2023
·
IF 9.8
Fang Deng,Jiafeng Pan,Manjia Chen,Zhi Liu,Junhua Chen,Chengshuai Liu
Polychlorinated biphenyls (PCBs) are ubiquitous persistent organic pollutants that cause harmful effects on environmental safety and human health. There is an urgent need to develop an intelligent method for PCBs sensing. In this work, we proposed a logic gate biosensing platform for simultaneous detection of multiple PCBs. 2,3′,5,5′-tetrachlorobiphenyl (PCB72) and 3,3′,4,4′-tetrachlorobiphenyl (PCB77) were used as the two inputs to construct biocomputing logic gates. We used 0 and 1 to encode the inputs and outputs. The aptamer was used to recognize the inputs and release the trigger DNA. A catalytic hairpin assembly (CHA) module is designed to convert and amplify each trigger DNA into multiple programmable DNA duplexes, which initiate the trans -cleavage activity of CRISPR/Cas12a for the signal output. The activated Cas12 cleaves the BHQ-Cy5 modified single-stranded DNA (ssDNA) to yield the fluorescence reporting signals. In the YES logic gate, PCB72 was used as the only input to carry out the logic operation. In the OR, AND, and INHIBIT logic gates, PCB72 and PCB77 were used as the two inputs. The output signals can be visualized by the naked eye under UV light transilluminators or quantified by a microplate reader. Our constructed biosensing platform possesses the merits of multiple combinations of inputs, intuitive digital output, and high flexibility and scalability, which holds great promise for the intelligent detection of different PCBs.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Hua Wang,Yang Sun,Yuan Zhou,Yujie Liu,Shuo Chen,Wenbo Sun,Zidong Zhang,Junqing Guo,Cheng Yang,Zhengping Li,Lei Chen
At present, the 100% case fatality and the cross-infection of virus strains make the ASFV 's harm to society continue to expand. The absence of an effective commercial vaccine poses early detection remains the most effective means of curbing ASFV infection. Here, we report a cascaded detection platform based on the CRISPR-Cas12a system combined with graphene field-effect transistor sensors. The cascade platform could detect ASFV as low as 0.5 aM within 30 min and achieve typing of wild and vaccine strains of ASFV in a single detection system. The evaluation of 16 clinical samples proved that, compared with the gold standard Real-time PCR method, this platform has outstanding advantages in sensitivity, specificity and typing. Combining CRISPR-Cas12a's high specificity with the bipolar electric field effect of graphene field-effect transistor, the cascade platform is expected to achieve clinical application in the field of DNA disease detection, and provides a new direction for multi-strain disease typing.
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高影响因子
JOURNAL OF CONTROLLED RELEASE
·
2020
·
IF 7.727
Xueqin Gao,Zhuang Jin,Xiangyu Tan,Chong Zhang,Chenming Zou,Wei Zhang,Jiahui Ding,Bhudev C. Das,Konstantin Severinov,Inga Isabel Hitzeroth,Priya Ranjan Debata,Dan He,Xin Ma,Xun Tian,Qinglei Gao,Jun Wu,Rui Tian,Zifeng Cui,Weiwen Fan,Zhaoyue Huang,Chen Cao,Yuxian Bao,Songwei Tan,Zheng Hu
Persistent high-risk HPV infection is the main factor for cervical cancer . HPV E7 oncogene plays an important role in HPV carcinogenesis. Down-regulation of E7 oncogene expression could induce growth inhibition in HPV-positive cells and thus treats HPV related cervical cancer. Here we developed a non-virus gene vector based on poly(amide-amine)-poly(β-amino ester) hyperbranched copolymer (hPPC) for the delivery of CRISPR/Cas9 system to specifically cleave HPV E7 oncogene in HPV-positive cervical cancer cells. The diameter of polyplex nanoparticles (NPs) formed by hPPCs/linear poly(β-amino ester) (PBAE) and plasmids were approximately 300 nm . These hPPCs/PBAE-green fluorescence protein plasmids polyplex NPs showed high transfection efficiency and low toxicity in cells and mouse organs. By cleaving HPV16 E7 oncogene, reducing the expression of HPV16 E7 protein and increasing intracellular retinoblastoma 1 (RB1) amount, hPPCs/PBAE-CRISPR/Cas9 therapeutic plasmids polyplex NPs, especially highly branched hPPC1-plasmids polyplex NPs, exhibited strong growth inhibition of cervical cancer cells in vitro and xenograft tumors in nude mice . Together, the hPPCs/PBAE polyplex NPs to deliver HPV16 E7 targeted CRISPR/Cas9 system in this study could potentially be applied to treat HPV-related cervical cancer.
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高影响因子
ACS Applied Materials & Interfaces
·
2025
·
IF 8.2
Kaiyong Yang,Chunli Zhang,Zeyu Wang,Qiqing Huang,Jing Qian,Gaoyu Shi,Wenwen Sun,Jinqiu Wang,Yu Ji,Zhaorui Sun,Yanni Song,Xin Han
EGFR tyrosine kinase inhibitors (EGFR-TKIs) have garnered substantial clinical success in treating nonsmall cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations. However, the inevitable emergence of drug resistance, frequently attributed to activation, mutation, or deletion of multiple signaling pathways, poses a significant challenge. Notably, the loss of PTEN protein expression has emerged as a pivotal mechanism fostering resistance in EGFR mutant lung cancers. Consequently, strategies aimed at upregulating PTEN expression hold great promise for restoring drug sensitivity. Leveraging the versatility, precision, and efficacy of nuclease-deactivated Cas9 (dCas9) as a transcriptional activator, we designed a CRISPR-dCas9 system to stimulate PTEN expression. To further enhance target specificity and drug delivery efficiency, we innovatively harnessed the tumor cell membrane (CCM) as a homologous targeting surface coating for our vector, thereby creating a targeted activation nanoplatform. Comprehensive in vitro and in vivo evaluations demonstrated that the synergistic interplay between gefitinib and the CRISPR-dCas9 system significantly enhanced drug sensitivity. The finding underscores the potential of our approach in addressing the issue of lung cancer resistance, offering a promising avenue for personalized and effective cancer therapies.
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高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Xiaotian Guan,Jingru Zhao,Zhou Sha,Yujie Liang,Jianghong Huang,Jun Zhang,Shuqing Sun
Exosomes, as novel biomarker for liquid biopsy, exhibit huge important potential value for cancer diagnosis. However, various proteins show different expression levels on exosomal membrane, and the absolute concentration of exosomes in clinical samples is easily influenced by a number of factors. Here, we developed a CRISPR/Cas12a and aptamer-chemiluminescence based analysis (CACBA) for the relative abundance determination of tumor-related protein positive exosomes in plasma for breast cancer diagnosis. The total concentration of exosomes was determined through captured CD63 using a CRISPR/Cas12a-based method with the LoD of 8.97 × 10 3 particles/μl. Meanwhile, EpCAM and MUC1 positive exosomes were quantitatively detected by aptamer-chemiluminescence (ACL) based method with the LoD of 1.45 × 10 2 and 3.73 × 10 2 particles/μl, respectively. It showed that the percentages of EpCAM and MUC1 positive exosomes offered an excellent capability to differentiate breast cancer patients and healthy donors. The high sensitivity, strong specificity, outstanding anti-interference capability, and steady recovery rate of this approach offered higher accuracy and robustness than the commercialized method in clinical trial. In addition with good stability, easy preparation and low cost, this method not only provides a new approach to rapid analysis of exosome proteins, it may be quickly extended to the diagnoses of various cancers.
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高影响因子
ACS Nano
·
2024
·
IF 15.8
Yu Ji,Suchen Qu,Gaoyu Shi,Liansheng Fan,Jing Qian,Zhaorui Sun,Feng Lu,Xin Han
Hypoxia is one of the most typical features among various types of solid tumors, which creates an immunosuppressive tumor microenvironment (TME) and limits the efficacy of cancer treatment. Alleviating hypoxia becomes a key strategy to reshape hypoxic TME which improves cancer immunotherapy. However, it remains challenging to perform tumor precision therapy with controllable switches through hypoxia-activated gene editing and prodrugs to alleviate hypoxia. In this study, silica-coated second near-infrared window (NIR-II) emitting silver sulfide quantum dots are used as the carrier to load the Clustered Regularly Interspaced Short Palindromic Repeats/Cas9 (CRISPR/Cas9) system to target hypoxia-inducible factor-1 (HIF-1α) and guide tumor-targeted imaging. To reduce the off-target effects in nontumor cells and better control safety risks, a TME-triggered cascade-activation nanodiagnostic and therapeutic platform (AA@Cas-H@HTS) is designed, which achieves the hypoxia activation of prodrug tirapazamine (TPZ) and spatiotemporal release of CRISPR/Cas9 ribonucleoprotein. Tumor hypoxia is greatly alleviated by the synergistic function of HIF-1α depletion by gene editing and TPZ activation. Importantly, targeting HIF-1α disrupts the programmed cell death 1/programmed cell death ligand 1 (PD-1/PD-L1) signaling pathway, which effectively reshapes the immune-suppressive TME and activates T cell-mediated antitumor immunity. Taken together, we have provided a TME-triggered cascade-activation nanoplatform to alleviate hypoxia for improved cancer immunotherapy.
研究领域:
癌症生物学
·
免疫学
·
纳米技术
·
分子生物学
·
诊疗学
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PubMed
高影响因子
Pharmaceutics
·
2024
·Vol. 16
·期 2
·213
·
IF 4.9
Jiahui Ding,Handan Zhang,Tianli Dai,Xueqin Gao,Zhongyuan Yin,Qiong Wang,Mengqi Long,Songwei Tan
Poly (β-amino ester) (PBAE) is an exceptional non-viral vector that is widely used in gene delivery, owing to its exceptional biocompatibility, easy synthesis, and cost-effectiveness. However, it carries a high surface positive charge that may cause cytotoxicity. Therefore, hydrophilic d-α-tocopherol polyethylene glycol succinate (TPGS) was copolymerised with PBAE to increase the biocompatibility and to decrease the potential cytotoxicity of the cationic polymer-DNA plasmid polyplex nanoparticles (NPs) formed through electrostatic forces between the polymer and DNA. TPGS-b-PBAE (TBP) copolymers with varying feeding molar ratios were synthesised to obtain products of different molecular weights. Their gene transfection efficiency was subsequently evaluated in HEK 293T cells using green fluorescent protein plasmid (GFP) as the model because free GFP is unable to easily pass through the cell membrane and then express as a protein. The particle size, ζ-potential, and morphology of the TBP2-GFP polyplex NPs were characterised, and plasmid incorporation was confirmed through gel retardation assays. The TBP2-GFP polyplex NPs effectively transfected multiple cells with low cytotoxicity, including HEK 293T, HeLa, Me180, SiHa, SCC-7 and C666-1 cells. We constructed aMUC2(Mucin2)-targeting CRISPR/cas9 gene editing system in HEK 293T cells, with gene disruption supported by oligodeoxynucleotide (ODN) insertion in vitro. Additionally, we developed anLMP1(latent membrane protein 1)-targeting CRISPR/cas9 gene editing system in LMP1-overexpressing SCC7 cells, which was designed to cleave fragments expressing the LMP1 protein (related to Epstein–Barr virus infection) and thus to inhibit the growth of the cells in vivo. As evidenced by in vitro and in vivo experiments, this system has great potential for gene therapy applications.Keywords:poly (β-amino ester);gene transfection;D-α-tocopherol polyethylene glycol succinate;copolymer;CRISPR/cas9 gene editing;gene therapy
研究领域:
生物技术
·
基因治疗
·
药学
·
分子生物学
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PubMed
高影响因子
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