LANGMUIR
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2024
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IF 3.7
Yaqin Zhang,Jianai Chen,Fange Kong,Chunxia Wang,Hangyu Guo,Yingchun Li,Jiahui Lu,Jicheng Zhang,Jiasi Wang,Yulin Zhou
Viral infections, such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are some of the most dangerous threats to humans. SARS-CoV-2 has caused a global pandemic, highlighting the unprecedented demand for rapid and portable diagnostic methods. To meet these requirements, we designed a label-free colorimetric platform that combines the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated proteins (Cas) 12a system for naked-eye detection (named LFP). This method utilizes reverse transcription loop-mediated isothermal amplification (RT-LAMP) and the trans-cleavage activity of the CRISPR/Cas12a system to increase the sensitivity and specificity of the reaction. This platform can detect as few as 4 copies/μL of RNA and produces no false positive results when tested against the influenza virus. To better meet the requirements of point-of-care (POC) detection, we developed a portable device that can be applied in resource-poor and densely populated regions. The LFP assay holds great potential for application in resource-limited settings, and the label-free gold nanoparticle (AuNPs) probe can reduce costs, making it suitable for large-scale screening. We expect that the LFP assay will be promising for the POC screening of COVID-19.
研究领域:
分子生物学
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生物技术
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病毒学
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纳米技术
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诊断方法
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PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2026
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IF 11.8
Lingsong Lu,Yingying Zhang,Bei Liu,Nuo zhou,Dao jun Yu,Ying Wang
Circular RNAs (circRNAs) represent an emerging family of noncoding transcripts defined by closed-loop architecture, which are now established as key participants in the etiology of tumorigenesis and malignant progression. While circRNAs show potential as therapeutic targets and biomarkers, the accurate detection of circRNAs remains challenging due to interference from homologous linear RNAs. In this study, an ultrasensitive method for detecting circ_0051240 based on the sulfhydrated toehold capture probe-initiator assembly-mediated hybridization chain reaction (HCR) and the CRISPR-Cas12a system was developed. The capture probe (with a toehold domain)-initiator strand duplex was ingeniously fabricated to identify the back-splice junction (BSJ) of circRNA. After magnetic enrichment and separation by the Fe 3 O 4 -Au@UIO-66-NH 2 nanocomposite, the initiator strand (H0) was delivered to trigger HCR. The HCR product dsDNA concatemers contained multiply repeated CRISPR-targetable DNA sites that were readily recognized by the CRISPR RNA (crRNA). This specific recognition and binding activated the CRISPR-Cas12a system's collateral endonuclease activity, leading to cleavage of the fluorophore-quencher (FQ) reporters and fluorescence emission at a characteristic wavelength. This design eliminated linear RNA-related interference and enhanced the detection of fluorescence intensity (FI). Under optimal conditions, the proposed HCR/CRISPR-Cas12a method exhibited a wide quantitative measurement range spanning from 45 pM up to 180 nM, achieving a notable limit of detection (LOD) of 0.03 pM. In this study, a novel circRNA sensing strategy capable of accurate and highly sensitive quantification of ovarian cancer-specific circRNA was reported. The proposed method exhibits acceptable performance when compared to present approaches.
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高影响因子
Environmental Science-Nano
·
2022
·Vol. 9
·期 9
·3417-3426
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IF 9.473
Fan Yang,Wei Wang,Mei Zhang,Wenxin Tao,Youwang Wang,Jiameng Shi,Yuedi Ding,Minhao Xie,Sai Zhang,Zhenqiang Fan,Kai Zhang
Since the emergence of coronavirus disease 2019 (COVID-19), this highly contagious disease has ravaged the world, cumulatively causing millions of deaths and huge economic losses. As the culprit of COVID-19, severe acute respiratory syndrome β-coronavirus 2 (SARS-CoV-2) is highly infectious and pathogenic, which has caused extreme panic worldwide. Early and rapid monitoring of SARS-CoV-2 has a significant role in suppressing the pandemic and reducing the virus's lethality. In our work, we prepared a self-enhanced ruthenium complex linked to zeolitic imidazole framework-8 (ZIF-8) and used it as an electrochemiluminescence (ECL) emitter. Additionally, a double-stranded specific nuclease (DSN)-assisted target RNA cycling with catalytic hairpin assembly (CHA) signal amplification technology was used to achieve the conversion of target RNA concentration to double-stranded DNA (dsDNA) output which significantly improved the detection sensitivity of target RNA under environmental conditions and in real human serum samples. In addition, we also combined the trans-cleavage property of CRISPR–Cas12a with the adsorption property of C3N4 on a ferrocene (Fc)-labeled DNA probe and obtained target RNA-dependent ECL signals. The reliable detection protocol achieved the transformation of SARS-CoV-2 RNA concentration to ECL responses, obtaining a limit of detection (LOD) of 0.67 fM with high specificity and reproducibility, which was of guiding significance for current detection methods of mutant SARS-CoV-2 and universal RNA.
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高影响因子
FOOD CONTROL
·
2025
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IF 6.3
Xinru Ren,Weiqing Sun,Bowen Li,Yijing Xiao,Xudong Yue,Meili Yang,Qing-ao Pang,Rui Zhu,Zhiqiang Guo,Haiping Zhang,Yu Wang,Su Liu,Jiadong Huang
The detection of pathogenic bacteria plays a crucial role in ensuring food safety and public health. The integration of isothermal amplification technology with the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) system offers a versatile approach to detecting foodborne pathogens. However, this method is often constrained by the incompatibility of the reaction systems and the lack of control over CRISPR/Cas activity. Here, we demonstrated a fluorescence biosensing strategy based on linear causal-regulated and autocatalysis-driven DNA circuits (termed LC@ADC) for highly sensitive and specific detection of Salmonella Typhimurium ( S. Typhimurium ). The approach of combining hybridization chain reaction (HCR) with DNAzyme-driven CRISPR/Cas12a activation was proposed for the first time, enabling not only one-step cascade signal amplification but also controllable DNA circuits. The LC@ADC strategy is capable of quantifying concentrations of S. Typhimurium ranging from 10 to 1 × 10 5 CFU/mL, with a detection limit as low as 6.53 CFU/mL. Furthermore, the LC@ADC strategy has been successfully employed for the detection of S. Typhimurium in samples of orange juice, chicken, and lake water. Therefore, this strategy provides a novel approach for the high-performance detection of pathogenic bacteria.
研究领域:
分子生物学
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生物技术
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食品安全
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公共卫生
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生物传感器
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微生物学
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高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
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IF 13.3
Haoyu Wang,Shiting Chen,Jiqing Liu,Wanyue Liu,Wei Chen,Lin Huang,Chao Yan
In recent years, RPA-CRISPR/Cas12a-based electrochemical biosensors have emerged as highly effective tools for nucleic acid detection. However, the random trans -cleavage characteristics of Cas12a on single-stranded nucleic acid sequences can result in the presence of uncertain nucleic acid residues on the electrode surface post-reaction. This situation complicates subsequent sensitization strategies. In this study, we devised a universal sensitization protocol mediated by terminal deoxynucleotidyl transferase (TdT) , a template-independent DNA polymerase and G-rich spherical nucleic acids (G-rich SNAs) with superior performance. Initially, Cas12a was activated by the specific RPA amplification product and cleaved the closed single stranded DNA (C-ssDNA) on the electrode surface. Then, the TdT enzyme extended residual single stranded DNA (R-ssDNA) on the electrode surface, forming G-Quadruplex structures. Subsequently, G-rich SNAs were immobilized on the electrode surface via π-π stacking interactions with the G-quadruplexes. Consequently, a substantial accumulation of hemin occurred within the G-quadruplex cluster structures, generating significant electrochemical signals. We applied this method to the trace detection of Staphylococcus aureus ( S. aureus ) and achieved a good linear calibration curve with concentrations ranging from 0 CFU/mL to 10 4 CFU/mL. Notably, the limit of detection (LOD) was as low as 1.29 CFU/mL, surpassing that of most similar sensors. These results demonstrate that our proposed universal sensitization method effectively addresses the limitations associated with the random trans -cleavage characteristics of Cas12a.
研究领域:
生物传感
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纳米技术
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分子生物学
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电化学
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高影响因子
ACS Applied Materials & Interfaces
·
2024
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IF 8.3
Wenyue Zhang,Ziyi Li,Xiaoli Lun,Yingshu Guo
Targeting tumor markers is one of the most important approaches to tumor therapy, and the “suicide” pattern of tumor marker response is a very challenging study. Telomerase, as one of the key factors associated with human longevity and cancer progression, is considered to be an emerging biomarker for cancer diagnosis. The targeted drug delivery nanobomb─BIBR1532@HSN/FQDNA/MUC1 aptamer (B@HDA) is prepared in this study based on hollow silica nanoparticles (HSN) and CRISPR systems. Amino-modified FQDNA and amino-modified MUC1 aptamer are covalently attached to the surface of carboxyl-functionalized HSN. The modified MUC1 aptamer directs the nanobomb to specifically target breast cancer cells (MCF-7) and FQDNA sequesters the telomerase inhibitor (BIBR1532) within the HSN. Telomerase primers (TPs) is recognized by the highly expressed telomerase in MCF-7 cells and is elongated to form DNA substrates. The substrate pairs with crRNA bases to effectively activate CRISPR-Cas12a. The activated CRISPR-Cas12a precisely cut FQDNA, releasing BIBR1532, which inhibits telomerase activity. This strategy achieves telomerase “suicide”. The nanobomb described above has the following advantages. (1) The “closing” effect of FQDNA contributes to reducing the nonspecific release of BIBR1532. (2) B@HDA, combined with CRISPR, regulates mitochondrial dysfunction and cell senescence in MCF-7 cells. (3) In the tumor-bearing mouse model, B@HDA, combined with CRISPR, exhibits good biocompatibility and an obvious tumor ablation effect on MCF-7 tumors, suggesting potential application prospects across a wide range of cancer cell lines. In summary, the proposed nanobomb provides a tunable switch approach for the specific inhibition of telomerase and the reduction of tumor cell growth, representing a promising avenue for promoting senescence and treating cancer.
研究领域:
癌症研究
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分子生物学
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纳米技术
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基因工程
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高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Peiqing Luo,Xiaocui Huang,Fang Luo,Zhonghui Chen,Yu Chen,Cuiying Lin,Jian Wang,Bin Qiu,Zhenyu Lin
In this work, combined with the high amplification efficiency of hybridization chain reaction (HCR), high specificity of the CRISPR/Cas12a system, and convenience of the homogeneous electrochemiluminescence (ECL) assay based on the regulation of negative charge on the reporting probes, a sensitive ECL biosensor for hepatitis B virus DNA (chosen as a model target) had been developed. The initiator chain trigger DNA that can induce HCR amplification is modified on the surface of ruthenium bipyridine-doped silica nanoparticles (Ru@SiO2 NPs) first, and large amounts of negative charges modified on the particles were achieved through the HCR amplification reaction. The efficiency of the nanoparticles reaching the negatively charged working electrode can be regulated and realize the change of the ECL signal. In addition, long DNA on the surface of the luminescent body may prevent the coreactant from entering the pore to react with ruthenium bipyridine. These factors combine to produce a low-background system. The presence of the target can activate the CRISPR/Cas12a system and make trigger DNA disappear from the nanoparticle surface, and strong ECL can be detected. The sensor does not require a complex electrode modification; therefore, it has better reproducibility. Additionally, due to dual signal amplification, the sensor has a high sensitivity. In the range of 10 fM to 10 nM, the ECL intensity exhibits a strong linear relationship with the logarithm of the target concentration, and the detection limit is 7.41 fM. This sensor has shown high accuracy in detecting clinical samples, which holds significant potential for application in clinical testing.
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高影响因子
Environmental Science-Nano
·
2021
·Vol. 9
·期 1
·162-172
·
IF 8.131
Kai Zhang,Zhenqiang Fan,Yuedi Ding,Sha Zhu,Minhao Xie,Nan Hao
In this paper, we present the first idea of using a DNA triple helix structure to inhibit CRISPR-Cas12a activity and apply it to the design of an electrochemiluminescent biosensor for the detection of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA-dependent RNA polymerase (RdRp) gene in real samples and environmental surveillance. We employed a segment from the RdRp gene of SARS-CoV-2 by an entropy-driven reaction, which was paired with double-stranded DNA that can activate CRISPR-Cas12a activity by Hoogsteen pairing to form triple-stranded DNA, thereby inhibiting the binding interaction of the double-stranded DNA with CRISPR-Cas12a, which in turn inhibits the trans cleavage activity of CRISPR-Cas12a. The inhibited CRISPR-Cas12a is unable to cut the nucleic acid modified on the electrode surface, resulting in the inability of the ferrocene (Fc) modified on the other end of the nucleic acid to move away from the electrode surface, and thus failing to cause electrochemiluminescence changes in GOAu–Ru modified on the electrode surface. The extent of the electrogenic chemiluminescence change can reflect the concentration of the gene to be tested. Using this system, we achieved the detection of the SARS-CoV-2 RdRp gene with a detection limit of 32.80 aM.
研究领域:
分子生物学
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生物技术
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分析化学
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病毒学
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高影响因子
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