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一种将CRISPR-Cas12a与适体相结合的多功能生物传感平台,用于检测多种分析物。

A versatile biosensing platform coupling CRISPR-Cas12a and aptamers for detection of diverse analytes.

作者信息

Zhao Xiangxiang, Li Shanshan, Liu Guang, Wang Zhong, Yang Zhiheng, Zhang Quanwei, Liang Mindong, Liu Jiakun, Li Zilong, Tong Yaojun, Zhu Guoliang, Wang Xinye, Jiang Lan, Wang Weishan, Tan Gao-Yi, Zhang Lixin

机构信息

State Key Laboratory of Bioreactor Engineering, and School of Biotechnology, East China University of Science and Technology (ECUST), Shanghai 200237, China.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.

出版信息

Sci Bull (Beijing). 2021 Jan 15;66(1):69-77. doi: 10.1016/j.scib.2020.09.004. Epub 2020 Sep 4.

Abstract

Rapid and sensitive detection of various analytes is in high demand. Apart from its application in genome editing, CRISPR-Cas also shows promises in nucleic acid detection applications. To further exploit the potential of CRISPR-Cas for detection of diverse analytes, we present a versatile biosensing platform that couples the excellent affinity of aptamers for broad-range analytes with the collateral single-strand DNA cleavage activity of CRISPR-Cas12a. We demonstrated that the biosensors developed by this platform can be used to detect protein and small molecule in human serum with a complicated background, i.e., the tumor marker alpha fetoprotein and cocaine with the detection limits of 0.07 fmol/L and 0.34 μmol/L, respectively, highlighting the advantages of simplicity, sensitivity, short detection time, and low cost compared with the state-of-the-art biosensing approaches. Altogether, this biosensing platform with plug-and-play design show great potential in the detection of diverse analytes.

摘要

对各种分析物进行快速灵敏的检测需求很高。除了在基因组编辑中的应用外,CRISPR-Cas在核酸检测应用中也显示出前景。为了进一步挖掘CRISPR-Cas用于检测多种分析物的潜力,我们提出了一种通用的生物传感平台,该平台将适体对广泛分析物的优异亲和力与CRISPR-Cas12a的附带单链DNA切割活性相结合。我们证明,由该平台开发的生物传感器可用于检测背景复杂的人血清中的蛋白质和小分子,即肿瘤标志物甲胎蛋白和可卡因,检测限分别为0.07 fmol/L和0.34 μmol/L,与最先进的生物传感方法相比,突出了其简单、灵敏、检测时间短和成本低的优点。总之,这种具有即插即用设计的生物传感平台在检测多种分析物方面显示出巨大潜力。

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