Programmable Aptasensor for Regulating CRISPR/Cas12a Activity

文献类型： 外文期刊

第一作者： Yu, Yao

作者： Yu, Yao;Shi, Wen;Li, Qiaoyu;Shi, Wen;Yang, Yuxin;Dai, Junbiao;Mao, Guobin;Ma, Yingxin;Li, Qiaoyu;He, Hongpeng;Dai, Junbiao

作者机构：

关键词： aptamers; biosensors; CRISPR/Cas12a; molecular recognition; point-of-caretesting

期刊名称：ACS SENSORS （ 影响因子：8.9； 五年影响因子：9.0 ）

ISSN： 2379-3694

年卷期： 2023 年 9 卷 1 期

页码：

收录情况： SCI

摘要： CRISPR-mediated aptasensors have gained prevalence for detecting non-nucleic acid targets. However, there is an urgent need to develop an easily customizable design to improve the signal-to-noise ratio, enhance universality, and expand the detection range. In this article, we report a CRISPR-mediated programmable aptasensor (CPAS) platform. The platform includes single-stranded DNA comprising the aptamer sequence, locker DNA, and a crRNA recognition region, forming a hairpin structure through complementary hybridization. With T4 DNA polymerase, the crRNA recognition region was transformed into a complete double-stranded DNA through stem-loop extension, thereby activating the trans-cleavage activity of Cas 12a and generating fluorescence signals. The specific binding between the target molecule and aptamer disrupted the formation of the hairpin structure, altering the fluorescence signals. Notably, the CPAS platform allows for easy customization by simply changing the aptamer sequence and locker DNA, without entailing adjustments to the crRNA. The optimal number of bases in the locker DNA was determined to be seven nucleotides for the SARS-CoV-2 spike (S) protein and four nucleotides for ATP. The CPAS platform exhibited high sensitivity for S protein and ATP detection. Integration with a lateral flow assay enabled sensitive detection within 1 h, revealing its excellent potential for portable analysis.

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