CRISPR/Cas12a-loaded intelligent DNA hydrogel for universal and ultrasensitive exosome assay

IF 9.7 4区 医学 Q1 MATERIALS SCIENCE, BIOMATERIALS VIEW Pub Date : 2024-02-01 DOI:10.1002/viw.20230086
Jie Luo, Binpan Wang, Xiaoqi Tang, Ping Huang, Sha Yang, Shuang Zhao, Shuang Xie, Qiaofeng Li, Kai Chang, Ming Chen
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Abstract

Tumor-derived exosomes are crucial for early non-invasive and accurate tumor diagnosis in clinical diagnostics. The development of highly sensitive, simple, and intuitive exosome assays has sparked a research upsurge in clinical diagnostics. Here, we develop a bio-responsive intelligent DNA hydrogel loaded with CRISPR/Cas12a for universal and ultrasensitive detection of the exosomes. The aptamer serves as the target response unit and switch, competitively disintegrating the region of the DNA linkers and then Cas12a/crRNA was released and activated, resulting in a high fluorescent intensity for exosome detection at the detection limit of 119 particles/μL. Moreover, a constructed colorimetric tube is made by loading a colorimetric filter membrane on the tube lid and intelligent DNA hydrogel on the tube bottom, which enables one-pot portable colorimetric detection. Without the need for laboratory instruments and professionals, this strategy allows for naked eye detection with limit of detection as low as 104 particles/μL, and shows great applicability in distinguishing between healthy individuals, pretreatment patients, and post-treatment patients after obtaining a testable analyte. In this study, an ultrasensitive detection platform for exosomes that enables one-step sensing and dual signal output was introduced. The findings here suggest that this platform is a promising tool for the application of liquid biopsy based on exosomes in clinical diagnosis.

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用于通用超灵敏外泌体检测的 CRISPR/Cas12a 负载智能 DNA 水凝胶
在临床诊断中,肿瘤外泌体对于早期无创、准确的肿瘤诊断至关重要。高灵敏度、简单直观的外泌体检测方法的开发掀起了临床诊断研究的热潮。在这里,我们开发了一种载入 CRISPR/Cas12a 的生物响应式智能 DNA 水凝胶,用于外泌体的通用超灵敏检测。吻合器作为目标响应单元和开关,竞争性地分解 DNA 链接区域,然后释放并激活 Cas12a/crRNA,从而产生高荧光强度,用于外泌体检测,检测限为 119 个颗粒/μL。此外,通过在管盖上加载比色滤膜,在管底上加载智能 DNA 水凝胶,还制作了一个结构化比色管,实现了一次性便携式比色检测。这种策略无需实验室仪器和专业人员,可实现肉眼检测,检测限低至 104 粒子/μL,在获得可检测的分析物后,可用于区分健康人、治疗前病人和治疗后病人,显示出极大的适用性。本研究介绍了一种外泌体超灵敏检测平台,该平台可实现一步感应和双信号输出。研究结果表明,该平台是将基于外泌体的液体活检应用于临床诊断的理想工具。
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来源期刊
VIEW
VIEW Multiple-
CiteScore
12.60
自引率
2.30%
发文量
0
审稿时长
10 weeks
期刊介绍: View publishes scientific articles studying novel crucial contributions in the areas of Biomaterials and General Chemistry. View features original academic papers which go through peer review by experts in the given subject area.View encourages submissions from the research community where the priority will be on the originality and the practical impact of the reported research.
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