| 基于CRISPR/Cas技术的核酸检测研究进展 |
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| 引用本文： | 周桓,邵艳娜,王涓,吴清平,丁郁. 基于CRISPR/Cas技术的核酸检测研究进展[J]. 微生物学报, 2021, 61(12): 3856-3869 |
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| 作者姓名： | 周桓 邵艳娜 王涓 吴清平 丁郁 |
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| 作者单位： | 广东省科学院微生物研究所, 华南应用微生物国家重点实验室, 广东省微生物安全与健康重点实验室, 广东 广州 510070;暨南大学生命科学技术学院, 广东 广州 510632;广东省科学院微生物研究所, 华南应用微生物国家重点实验室, 广东省微生物安全与健康重点实验室, 广东 广州 510070;暨南大学食品科学与工程系, 广东 广州 510632;华南农业大学食品学院, 广东 广州 510640;广东省科学院微生物研究所, 华南应用微生物国家重点实验室, 广东省微生物安全与健康重点实验室, 广东 广州 510070;暨南大学生命科学技术学院, 广东 广州 510632;暨南大学食品科学与工程系, 广东 广州 510632 |
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| 基金项目： | 国家“食品安全重点研发”(2018YFC1602500)；广东省基础与应用基础研究重大项目(2020B0301030005)；国家自然科学基金(31730070)；广东省科学院重点科技计划(2019GDASYL-0201001)；广东省微生物安全与健康重点实验室基金(2020B121201009) |
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| 摘 要： | 由成簇、规则间隔的短回文重复序列（Clustered regularly interspaced short palindromic repeats，CRISPR）和CRISPR相关蛋白（CRISPR-associated protein，Cas）组成的CRISPR/Cas系统是广泛存在于多数细菌和古细菌中的一种适应性免疫系统。CRISPR/Cas系统可识别并结合外源入侵的核酸分子，之后Cas蛋白的切割活性被激活，能够对入侵的核酸分子进行切割使其降解。利用CRISPR/Cas系统特异的序列识别及切割活性，将其应用于核酸检测中，为提高检测灵敏度及特异性等性能指标提供了一种新思路。本文介绍了CRISPR/Cas系统的发展、作用机制等，对多样化的Cas蛋白在核酸检测中的代表性应用研究进行总结，进一步讨论了CRISPR/Cas技术应用于核酸检测中存在的优缺点，并对未来研究进行了展望，为基于CRISPR/Cas技术的核酸检测方法在病原微生物的检测中提供参考和依据。
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| 关 键 词： | CRISPR/Cas Cas蛋白 切割活性 核酸检测 病原微生物 |
| 收稿时间： | 2021-02-25 |
| 修稿时间： | 2021-05-14 |
Research progress of nucleic acid detection based on CRISPR/Cas technology |
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| Huan Zhou,Yanna Shao,Juan Wang,Qingping Wu,Yu Ding. Research progress of nucleic acid detection based on CRISPR/Cas technology[J]. Acta microbiologica Sinica, 2021, 61(12): 3856-3869 |
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| Authors: | Huan Zhou Yanna Shao Juan Wang Qingping Wu Yu Ding |
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| Affiliation: | Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, Guangdong Province, China;College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China;Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, Guangdong Province, China;Department of Food Science and Engineering, Jinan University, Guangzhou 510632, Guangdong Province, China;College of Food Science, South China Agricultural University, Guangzhou 510640, Guangdong Province, China; Guangdong Provincial Key Laboratory of Microbial Safety and Health, State Key Laboratory of Applied Microbiology Southern China, Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, Guangdong Province, China;College of Life Science and Technology, Jinan University, Guangzhou 510632, Guangdong Province, China;Department of Food Science and Engineering, Jinan University, Guangzhou 510632, Guangdong Province, China |
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| Abstract: | The CRISPR/Cas system, an adaptive immune system widely found in most bacteria and archaea, is composed of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas). The CRISPR/Cas system can recognize and bind foreign invading nucleic acid. Then the cleavage activity of Cas protein is activated that can cleave and degrade the invasive nucleic acid. The specific sequence recognition and cleavage activity of the CRISPR/Cas system is applied to nucleic acid detection that provides a new research idea to improve detection performance such as detection sensitivity and specificity. This study introduces the development of the CRISPR/Cas system, its mechanism of action, etc. In addition, we also summarize the representative applications of diversified Cas proteins in nucleic acid detection. The advantages and disadvantages of CRISPR/Cas technology in nucleic acid detection are discussed, and future research is proposed. In sum, this review aims to provide references for the detection of pathogenic microorganisms by using a nucleic acid detection method based on CRISPR/Cas technology. |
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| Keywords: | CRISPR/Cas Cas protein cleavage activity nucleic acid detection pathogenic microorganism |
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