基于四面体和 Cas12a 的双识别精确核酸探针揭示基因与环境的相互作用

IF 8 1区化学 Q1 CHEMISTRY, ANALYTICAL

Sensors and Actuators B: Chemical Pub Date : 2024-11-25 DOI:10.1016/j.snb.2024.137001

Linmin Zhong, Sirui Yi, Haiyan Qiu, Yan Yan, Fenying Kong, Shanwen Hu

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引用次数: 0

摘要

基因表达的精确检测对于理解生物与其环境之间的相互作用至关重要。在这项研究中，我们开发了一种基于CRISPR-Cas12a系统和DNA四面体的高灵敏度双识别基因检测方法，并在生物样本中进行了应用验证。通过优化 Cas12a 信号读出系统，该方法表现出优异的灵敏度和稳定性。进一步的实验表明，该方法能准确检测 LKB1 基因的表达，在 10 pM 至 10 nM 的浓度范围内线性关系良好，检测限低至 1 pM。我们利用砷诱导的小鼠肝损伤模型验证了该方法在复杂生物样本中的适用性，发现该检测系统在识别基因表达差异方面优于传统的 RT-qPCR，具有更高的灵敏度和特异性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

A dual-recognition precise nucleic acid probe based on tetrahedron and Cas12a reveals gene-environment interactions

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A dual-recognition precise nucleic acid probe based on tetrahedron and Cas12a reveals gene-environment interactions

Precise detection of gene expression is crucial for understanding the interactions between organisms and their environment. In this study, we developed a highly sensitive dual-recognition gene detection method based on the CRISPR-Cas12a system and DNA tetrahedron and validated its application in biological samples. By optimizing the Cas12a signal readout system, the method demonstrated excellent sensitivity and stability. Further experiments showed that the method could accurately detect the expression of the LKB1 gene, with good linearity observed in the concentration range of 10 pM to 10 nM, and a detection limit as low as 1 pM. Using an arsenic-induced mouse liver injury model, we validated the applicability in complex biological samples and found that this detection system outperformed traditional RT-qPCR in recognizing gene expression differences, offering higher sensitivity and specificity.

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来源期刊

Sensors and Actuators B: Chemical 工程技术-电化学

CiteScore

14.60

自引率

11.90%

发文量

1776

审稿时长

3.2 months

期刊介绍： Sensors & Actuators, B: Chemical is an international journal focused on the research and development of chemical transducers. It covers chemical sensors and biosensors, chemical actuators, and analytical microsystems. The journal is interdisciplinary, aiming to publish original works showcasing substantial advancements beyond the current state of the art in these fields, with practical applicability to solving meaningful analytical problems. Review articles are accepted by invitation from an Editor of the journal.