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基于适配体的CRISPR-Cas技术对多种生物标志物和小分子靶标的诊断

Aptamer-based CRISPR-Cas powered diagnostics of diverse biomarkers and small molecule targets.

作者信息

Kadam Ulhas Sopanrao, Cho Yuhan, Park Tae Yoon, Hong Jong Chan

机构信息

Division of Life Science and Division of Applied Life Science (BK21 Four), Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Jinju, Gyeongnam-do 52828 Republic of Korea.

Graduate School of Education, Yonsei University, Seoul, 03722 Republic of Korea.

出版信息

Appl Biol Chem. 2023;66(1):13. doi: 10.1186/s13765-023-00771-9. Epub 2023 Feb 18.

Abstract

CRISPR-Cas systems have been widely used in genome editing and transcriptional regulation. Recently, CRISPR-Cas effectors are adopted for biosensor construction due to its adjustable properties, such as simplicity of design, easy operation, collateral cleavage activity, and high biocompatibility. Aptamers' excellent sensitivity, specificity, in vitro synthesis, base-pairing, labeling, modification, and programmability has made them an attractive molecular recognition element for inclusion in CRISPR-Cas systems. Here, we review current advances in aptamer-based CRISPR-Cas sensors. We briefly discuss aptamers and the knowledge of Cas effector proteins, crRNA, reporter probes, analytes, and applications of target-specific aptamers. Next, we provide fabrication strategies, molecular binding, and detection using fluorescence, electrochemical, colorimetric, nanomaterials, Rayleigh, and Raman scattering. The application of CRISPR-Cas systems in aptamer-based sensing of a wide range of biomarkers (disease and pathogens) and toxic contaminants is growing. This review provides an update and offers novel insights into developing CRISPR-Cas-based sensors using ssDNA aptamers with high efficiency and specificity for point-of-care setting diagnostics.

摘要

CRISPR-Cas系统已广泛应用于基因组编辑和转录调控。最近,CRISPR-Cas效应器因其可调节的特性,如设计简单、操作容易、旁切活性和高生物相容性,而被用于构建生物传感器。适体具有出色的灵敏度、特异性、体外合成、碱基配对、标记、修饰和可编程性,使其成为CRISPR-Cas系统中极具吸引力的分子识别元件。在此,我们综述基于适体的CRISPR-Cas传感器的当前进展。我们简要讨论适体以及Cas效应蛋白、crRNA、报告探针、分析物的相关知识,以及靶标特异性适体的应用。接下来,我们介绍使用荧光、电化学、比色、纳米材料、瑞利和拉曼散射的制造策略、分子结合和检测方法。CRISPR-Cas系统在基于适体的多种生物标志物(疾病和病原体)和有毒污染物传感中的应用正在不断增加。本综述提供了最新信息,并为开发基于CRISPR-Cas的传感器提供了新的见解,这些传感器使用单链DNA适体,具有高效和特异性,可用于即时诊断。

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