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Molecular AND Logic Gate for Multiple Single-Nucleotide Mutations Detection Based on CRISPR/Cas9n System-Trigged Signal Amplification
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-05-01 , DOI: 10.1016/j.aca.2020.03.058
Weidan Chang 1 , Weipeng Liu 2 , Haoran Shen 2 , Sisi Chen 2 , Peizhen Liao 2 , Yingju Liu 3
Affiliation  

Precise detection of single-nucleotide mutations (SNMs) is extremely important in various biomedical applications, but the simultaneous detection of multiple SNMs remains a great challenge. Herein, we developed a new method based on CRISPR/Cas9 system for multiple SNMs detection. The CRISPR/Cas9 system transduces the nucleic acid into an intermediate trigger to initiate the isothermal amplification reaction and further form fluorescence signals. According to this strategy, we established nucleic acid bio-computing operations-molecule logic gate in simultaneous distinction of the genetic locus. We demonstrate that the fluorescence signals generated from different input combinations can be used to discriminate the multiple genetic locus, and the molecular logic gate has great potential in single-base mismatch detection. In addition, the successful assay of real samples indicates that the novel strategy could further adapt for the pathogenic monitoring and biomedical research.

中文翻译:

基于CRISPR/Cas9n系统触发信号放大的多单核苷酸突变检测的分子与逻辑门

单核苷酸突变 (SNM) 的精确检测在各种生物医学应用中极为重要,但同时检测多个 SNM 仍然是一个巨大的挑战。在此,我们开发了一种基于 CRISPR/Cas9 系统的多 SNM 检测新方法。CRISPR/Cas9系统将核酸转导为中间触发器,启动等温扩增反应,进一步形成荧光信号。根据这一策略,我们建立了核酸生物计算操作-分子逻辑门,同时区分了基因座。我们证明了由不同输入组合产生的荧光信号可用于区分多个基因位点,并且分子逻辑门在单碱基错配检测中具有巨大潜力。此外,
更新日期:2020-05-01
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