CRISPR-/Cas12a-Mediated Liposome-Amplified Strategy for the Surface-Enhanced Raman Scattering and Naked-Eye Detection of Nucleic Acid and Application to Food Authenticity Screening
CRISPR-/Cas12a-Mediated Liposome-Amplified Strategy for the Surface-Enhanced Raman Scattering and Naked-Eye Detection of Nucleic Acid and Application to Food Authenticity Screening
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DOI：
10.1021/acs.analchem.1c01163
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发表时间：
2021-07-19
影响因子：
7.4
通讯作者：
Li, Guoliang
中科院分区：
文献类型：
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作者：
Liu, Jianghua;Chen, Jiahui;Li, Guoliang
表面增强拉曼散射（SERS）由于其超高的灵敏度和独特的指纹信息，已被公认为生物传感器的有力工具。然而，由于信号转导和扩增策略的限制，痕量靶核酸检测存在一定的局限性。受具有特异性靶DNA激活侧支单链DNA （ssDNA）裂解活性的CRISPR/Cas12a和具有信号分子负载特性的脂质体的启发，我们首次提出了一种敏感的基于sers的现场核酸检测策略，该策略由具有反式裂解活性的CRISPR/Cas12a介导，用于捕获脂质体。负载4-硝基噻吩酚（4-NTP）和半胱氨酸两种信号分子的脂质体可以分别实现SERS和裸眼对靶DNA的双模检测。迅速放大的信号是由信号分子负载脂质体的触发分解引起的。解放的4-NTP，一个生物沉默拉曼报告，将实现高选择性和敏感的SERS测量。释放的半胱氨酸诱导等离子体金纳米粒子聚集，导致明显的红蓝比移，从而实现便携式裸眼检测。利用该策略，靶核酸浓度可以灵巧地转化为SERS和可视化信号，分别可以低至100am和10pm进行检测。该方法还成功应用于肉类掺假检测，实现了在复杂食品基质中低掺假率的检测。我们预计，该策略不仅将被视为食品真实性现场检测的通用平台，而且将扩大SERS的应用范围，以准确测定各种生物标志物。
Surface-enhanced Raman scattering (SERS) has been recognized as a powerful tool for biosensors due to the ultrahigh sensitivity and unique fingerprint information. However, there are some limitations in trace target nucleic acid detection for the restricted signal- transducing and amplification strategies. Inspired by CRISPR/Cas12a with specific target DNA-activated collateral single-strand DNA (ssDNA) cleavage activity and liposome with signal molecule-loading properties, we first proposed a sensitive SERS-based on-site nucleic acid detection strategy mediated by CRISPR/Cas12a with trans-cleavage activity on ssDNA linkers utilized to capture liposomes. Liposomes loading two kinds of signal molecules, 4-nitrothiophenol (4-NTP) and cysteine, could achieve the dual-mode detection of target DNA with SERS and naked eye, respectively. The promptly amplified signals were initiated by the triggered breakdown of signal molecule-loaded liposomes. Emancipated 4-NTP, a biological-silent Raman reporter, would achieve highly selective and sensitive SERS measurement. Released cysteine induced the aggregation of plasmonic gold nanoparticles, leading to an obvious red to blue colorimetric shift to realize portable naked-eye detection. With this strategy, target nucleic acid concentration was dexterously converted into SERS and visualization signals and could be detected as low as 100 aM and 10 pM, respectively. The approach was also successfully applied to determine meat adulteration, achieving the detection of a low adulteration ratio in the complicated food matrix. We anticipate that this strategy will not only be regarded as a universal platform for the on-site detection of food authenticity but also broaden SERS application for the accurate determination of diverse biomarkers.