Fluorescent aptasensor based on the MNPs-CRISPR/Cas12a-TdT for the determination of nasopharyngeal carcinoma-derived exosomes

Yi, Peng; Luo, Dan; Gao, Zhong Feng; Chen, Qinhua; Zhou, Yajuan

doi:10.1007/s00604-023-05657-7

Cited by 8 publications

(6 citation statements)

References 35 publications

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“…29 The schematic diagram is illustrated in Figure 2a. [29][30][31][32] Combining hybridization chain reaction (HCR) and CRISPR-Cas12a, the method achieved a low limit of detection (LOD) of 102 particles/µL, surpassing traditional assays like aptamer-ELISA and apta-HCR-ELISA. Application to clinical samples revealed diagnostic potential for nucleolin+ TEVs in nasopharyngeal carcinoma (NPC) and therapeutic monitoring using PD-L1+ TEVs.…”

Section: Detection Of Exosomesmentioning

confidence: 99%

“…Moreover, this approach enables the simultaneous detection of multiple targets in tumor exosomes and demonstrates the potential of liquid biopsy technology. Yi et al 31 reported a fluorescent aptamer sensor based on magnetic nanoparticles, CRISPR/Cas12a system, and terminal transferase for detecting exosomes from nasopharyngeal carcinoma. The illustrative diagram is presented in Figure 2c.…”

Section: Detection Of Exosomesmentioning

confidence: 99%

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Advances in CRISPR Based Biosensing Strategies for Cancer Diagnosis

Yang,

Zhong,

Zhong

et al. 2024

Cancer Screen Prev

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With the development of gene editing technology, its application in tumor diagnosis is becoming increasingly widespread. The CRISPR/Cas system is an important gene editing tool that can significantly improve the early detection rate and precision diagnosis level, enabling high-throughput and high-sensitivity detection of tumors. This article focuses on CRISPR/Cas system for detecting various tumor-related targets and elaborates on its applications in tumor diagnosis from five aspects: (1) detection of tumor-derived exosomes: by recognizing the surface proteins or nucleic acids of exosomes secreted by tumor cells into blood or other samples through adaptors, the CRISPR system is activated, achieving non-invasive liquid biopsy of tumors; (2) detection of circulating tumor DNA tumor cells disseminate DNA into the circulatory system to trigger nucleic acid reactions involving gene editing enzymes, enabling the monitoring of tumor dynamic states; (3) detection of circulating tumor cells (CTCs): by using aptamers to recognize surface proteins of tumor cells or directly detecting tumor-related nucleic acids, the integrated CRISPR system allows for the detection of circulating tumor cells even in trace amounts, achieving precise diagnosis; (4) detection of tumor markers: high sensitivity is achieved through the coupling of various tumor marker aptamers and gene editing systems; (5) detection and identification of tumor microenvironments: by activating gene editing enzyme activity through differential factors in the tumor tissue microenvironment and triggering nucleic acid reactions, the diagnosis and dynamic monitoring of tumors can be achieved. The progress and bottlenecks of the CRISPR/Cas system in tumor diagnosis in the future are also discussed.

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Section: Detection Of Exosomesmentioning

confidence: 99%

Section: Detection Of Exosomesmentioning

confidence: 99%

Advances in CRISPR Based Biosensing Strategies for Cancer Diagnosis

Yang,

Zhong,

Zhong

et al. 2024

Cancer Screen Prev

View full text Add to dashboard Cite

show abstract

“…25 Additionally, in fluorescence-based aptasensors, the instability of fluorescent substances often limits the detection sensitivity. 26 So far, there has been a need for novel aptamer sensor methods for effective detection of exosomes, combining the advantages of low electrochemical analysis detection limits and rapid fluorescence analysis to promote clinical diagnostics. Here, carbon and nitrogen nanosheets (g-C 3 N 4 ) 27 are combined with cobaltdoped single-atom nanoenzymes (Co-SANs) 28 and electrochemical-label prussian blue nanoparticles (PB) 29 followed by binding with single-stranded DNA aptamers and gold nanoparticles, to form a sensing platform (Scheme 1).…”

Section: Introductionmentioning

confidence: 99%

“…However, electrochemical aptasensors require complex and repetitive electrode incubations, leading to significantly lengthy processing times before detecting exosomes . Additionally, in fluorescence-based aptasensors, the instability of fluorescent substances often limits the detection sensitivity . So far, there has been a need for novel aptamer sensor methods for effective detection of exosomes, combining the advantages of low electrochemical analysis detection limits and rapid fluorescence analysis to promote clinical diagnostics.…”

Section: Introductionmentioning

confidence: 99%

Dual-Modal Aptasensor for Sensitive Detection of Non-Small Cell Lung Cancer Exosomes Utilizing Two-Dimensional Nanopaper Co@g-C₃N₄@PB

Xu,

Zhang,

et al. 2024

ACS Omega

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Nonsmall cell lung cancer (NSCLC), due to its lack of early symptoms, has become one of the leading causes of cancerrelated deaths globally. Exosomes, small membrane vesicles secreted by cells, are widely present in human bodily fluids. In the bodily fluids of NSCLC patients, the quantity of extracellular vesicles is double that of healthy individuals, suggesting their potential as biomarkers for screening NSCLC. This study designed a dual-modal aptasensor that integrated excellent sensitivity in electrochemical detection and portability in fluorescence detection into one device. AuNPs were functionalized with exosomecapturing probes containing thiol-modified CD63 aptamers, which were immobilized on screen-printed gold electrodes. On the other hand, the carboxylated CD63 aptamer was immobilized on the surface of PB-modified g-C 3 N 4 loaded with Co-SANs particles

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“…CRISPR-associated (CRISPR-Cas) effector protein system with nonspecific trans -cleavage activity enables amplified nucleic acid detection. , Among CRISPR/Cas family, the CRISPR/Cas12a system can cleave DNA labeled with a fluorophore-quencher (FQ) reporter and generate fluorescence signals for target detection. − Thus, the trans -cleavage ability of Cas12a activated by specific targets provides a versatile strategy to develop fluorescence biosensors. − However, the sensitivity of CRISPR/Cas12a system alone is insufficient to detect trace biomarkers in the early stage of diseases. , To improve its sensitivity, cascade amplification that combines CRISPR/Cas12a with other signal amplification methods is necessary. Thus, some CRISPR/Cas12a-based methods have been developed for tEV detection. − For example, Xing et al developed a dual amplification system based on HCR and CRISPR/Cas12a for nucleolin + or programmed death ligand 1+ (PD-L1+) tEV detection with a limit of detection (LOD: 10 2 particles/μL) through corresponding aptamer targeted recognition, respectively . Luo et al reported a platform for rapidly detecting PD-L1+ tEVs from nonsmall cell lung carcinoma.…”

mentioning

confidence: 99%

DNA Gate-Based CRISPR-Cas Exponential Amplification System for Ultrasensitive Small Extracellular Vesicle Detection to Enhance Breast Cancer Diagnosis

Tan,

Zhang,

Cheng

et al. 2024

Anal. Chem.

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Tumor-derived small extracellular vesicles (tEVs) as potential biomarkers possess abundant surface proteins closely related to parent cells, which are crucial for noninvasive cancer diagnosis. However, tEVs exhibit phenotype heterogeneity and low abundance, posing a significant challenge for multiplex detection with a high sensitivity. Herein, we developed a DNA gate-based exponential amplification CRISPR-Cas (DGEAC) system for accurate and ultrasensitive detection of tEVs, which can greatly improve the accuracy of breast cancer (BC) diagnosis. Based on the coexpression of CD63 and vascular endothelial growth factor (VEGF) on BC-derived tEVs, we developed a dual-aptamer-based AND gate fluorescent probe by proximity hybridization. By integrating the target recognition and trans-cleavage activity of Cas12a, an autocatalysis-driven exponential amplification circuit was developed for ultrasensitive detection of CD63 and VEGF proteins on tEVs, which could avoid false negative signals from single protein or other interfering proteins. We achieved highly sensitive detection of tEVs over a linear range from 1.75 × 10 3 to 3.5 × 10 8 particles/mL with a detection limit as low as 1.02 × 10 3 particles/mL. Furthermore, the DGEAC system can distinguish tEVs from tEVs derived from different BC cell lines, including MDA-MB-231, MCF-7, SKBR3, and MCF-10A. Compared to linear amplification (AUC 90.0%), the DGEAC system effectively differentiates BC in different stages (AUC 98.3%).

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Fluorescent aptasensor based on the MNPs-CRISPR/Cas12a-TdT for the determination of nasopharyngeal carcinoma-derived exosomes

Cited by 8 publications

References 35 publications

Advances in CRISPR Based Biosensing Strategies for Cancer Diagnosis

Advances in CRISPR Based Biosensing Strategies for Cancer Diagnosis

Dual-Modal Aptasensor for Sensitive Detection of Non-Small Cell Lung Cancer Exosomes Utilizing Two-Dimensional Nanopaper Co@g-C₃N₄@PB

DNA Gate-Based CRISPR-Cas Exponential Amplification System for Ultrasensitive Small Extracellular Vesicle Detection to Enhance Breast Cancer Diagnosis

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Fluorescent aptasensor based on the MNPs-CRISPR/Cas12a-TdT for the determination of nasopharyngeal carcinoma-derived exosomes

Cited by 8 publications

References 35 publications

Advances in CRISPR Based Biosensing Strategies for Cancer Diagnosis

Advances in CRISPR Based Biosensing Strategies for Cancer Diagnosis

Dual-Modal Aptasensor for Sensitive Detection of Non-Small Cell Lung Cancer Exosomes Utilizing Two-Dimensional Nanopaper Co@g-C3N4@PB

DNA Gate-Based CRISPR-Cas Exponential Amplification System for Ultrasensitive Small Extracellular Vesicle Detection to Enhance Breast Cancer Diagnosis

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Product

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Dual-Modal Aptasensor for Sensitive Detection of Non-Small Cell Lung Cancer Exosomes Utilizing Two-Dimensional Nanopaper Co@g-C₃N₄@PB