清脆的
适体
核酸
生物传感器
纳米技术
核酸检测
线性范围
DNA微阵列
核酸定量
检出限
计算机科学
计算生物学
化学
材料科学
分子生物学
生物
色谱法
生物化学
基因
基因表达
作者
Long Ma,Dan Liao,Zhiying Zhao,Jun Kou,Haoyu Guo,Xin Xiong,Shuli Man
出处
期刊:ACS Sensors
[American Chemical Society]
日期:2023-01-18
卷期号:8 (3): 1076-1084
被引量:32
标识
DOI:10.1021/acssensors.2c02097
摘要
Next-generation biosensing tools based on CRISPR/Cas have revolutionized the molecular detection. A number of CRISPR/Cas-based biosensors have been reported for the detection of nucleic acid targets. The establishment of efficient methods for non-nucleic acid target detection would further broaden the scope of this technique, but up to now, the concerning research is limited. In the current study, we reported a versatile biosensing platform for non-nucleic acid small-molecule detection called SMART-Cas12a (small-molecule aptamer regulated test using CRISPR/Cas12a). Simply, hybridization chain reaction cascade signal amplification was first trigged by functional nucleic acid (aptamer) through target binding. Then, the CRISPR/Cas system was integrated to recognize the amplified products followed by activation of the trans-cleavage. As such, the target can be ingeniously converted to nucleic acid signals and then fluorescent signals that can be readily visualized and analyzed by a customized 3D-printed visualizer with the help of a home-made App-enabled smartphone. Adenosine triphosphate was selected as a model target, and under the optimized conditions, we achieved fine analytical performance with a linear range from 0.1 to 750 μM and a detection limit of 1.0 nM. The satisfactory selectivity and recoveries that we have obtained further demonstrated this method to be suitable for a complex sample environment. The sample-to-answer time was less than 100 min. Our work not only expanded the reach of the CRISPR-Cas system in biosensing but also provided a prototype method that can be generalized for detecting a wider range of analytes with desirable adaptability, sensitivity, specificity, and on-site capability.
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