利用基于 COF-AuNPs 传感平台的 CRISPR/Cas9 nickase 驱动的 3D DNA walker 超灵敏检测循环肿瘤 DNA

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL Microchimica Acta Pub Date : 2024-10-15 DOI:10.1007/s00604-024-06749-8

Yuxin Wei, Yang Fu, Chuanhai Li, Siyu Chen, Linzhi Xie, Mei Chen

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引用次数: 0

摘要

我们设计了一种电化学生物传感器，利用 CRISPR Cas9n 驱动的 DNA 走行器与类金纳米共价有机框架（COFs-AuNPs）相结合来检测乳腺癌标志物（PIK3CA E545K ctDNA）。DNA walker 探针只有在存在循环肿瘤脱氧核糖核酸（ctDNA）时才会被激活，与支持探针结合形成双链，然后被 Cas9n/sgRNA 复合物特异性地裂解。这种裂解产生大量 DNA 片段，用于信号放大。作为电极材料的 COF-AuNPs 可促进电子转移，并为固定核酸探针提供额外的活性位点。该装置的检测限高达 1.76 aM，显示出极高的灵敏度。此外，Cas9n 还能提高传感器的特异性，准确区分一对碱基不匹配的序列，减少假阳性的出现。总之，该传感器具有出色的选择性和可重复性，有望用于乳腺癌的早期诊断。

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Ultrasensitive detection of circulating tumor DNA using a CRISPR/Cas9 nickase-driven 3D DNA walker based on a COF-AuNPs sensing platform

A electrochemical biosensor was designed utilizing a CRISPR Cas9n-driven DNA walker combined with gold-nanosphere-like covalent organic frameworks (COFs-AuNPs) to detect breast cancer markers (PIK3CA E545K ctDNA). The DNA walker probe is activated only in the presence of circulating tumor deoxyribonucleic acid (ctDNA), binding to a support probe to form a double strand that is then specifically cleaved by the Cas9n/sgRNA complex. This cleavage produces numerous DNA fragments for signal amplification. The COF-AuNPs as electrode materials facilitate electronic transfer and provide additional active sites for the immobilization of nucleic acid probes. This setup achieves a detection limit of 1.76 aM, demonstrating high sensitivity. Additionally, Cas9n improves the specificity of the sensor, accurately distinguishing a pair of base-mismatched sequences, and reducing the occurrence of false positives. Overall, the sensor exhibits excellent selectivity, reproducibility, and potential for early diagnosis of breast cancer.

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.