CRISPR-Cas系统在核酸检测中的应用研究

doi:10.13560/j.cnki.biotech.bull.1985.2021-0015

摘要/Abstract

摘要：

CRISPR-Cas系统是一种高效、实用的基因编辑工具,被广泛应用于基因组编辑和调控机制研究。目前发现的Cas系列核酸酶工具为开发用于各种目的的不同类型核酸检测,具有广阔的应用前景。CRISPR-Cas系统的高灵敏度和高特异性,使其在病原体检测、单核苷酸多态性（SNP）分析以及基因突变检测等发挥了极其重要的作用;另外,CRISPR-Cas系统在核酸检测领域的精确、高效性,间接推动了基础生物学和应用生物学研究的进展。概述了不同类型的靶向特定核酸检测的CRISPR-Cas系统的最新进展和用途,以及与之对应的新一代的体外检测平台,旨为核酸检测领域提供新的研究思路和理论依据。

关键词: 基因编辑, CRISPR-Cas系统, Cas蛋白核酸酶, 核酸检测

Abstract:

CRISPR-Cas is an efficient and practical gene editing tool,which is widely used in genome editing and regulation. Due to its high sensitivity and specificity,the CRISPR-Cas system also plays an important role in nucleic acid detection. It can detect pathogens,analyze single nucleotide polymorphisms（SNPs）,and detect genetic mutations. The currently discovered Cas series nuclease tools open the door to the development of new strategies for different types of nucleic acid detection for various purposes. In addition,the CRISPR-Cas system is used in nucleic acid detection. The accuracy and efficiency of the field indirectly promote the progress of basic biology and applied biology research. The article outlines the latest developments and uses of different types of CRISPR-Cas systems that target specific nucleic acid detection,and the corresponding new generation of in vitro detection platforms,which provide new research ideas and theoretical basis for the field of nucleic acid detection.

Key words: gene editing, CRISPR-Cas system, Cas protein nuclease, nucleic acid detection

胡秀文, 刘华, 王宇, 唐雪明, 王金斌, 曾海娟, 蒋玮, 李红. CRISPR-Cas系统在核酸检测中的应用研究[J]. 生物技术通报, 2021, 37(9): 266-273.

HU Xiu-wen, LIU Hua, WANG Yu, TANG Xue-ming, WANG Jin-bin, ZENG Hai-juan, JIANG Wei, LI Hong. Application of CRISPR-Cas System in Nucleic Acid Detection[J]. Biotechnology Bulletin, 2021, 37(9): 266-273.

图/表 1

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效应物 Effector	结构域 Domain	向导RNA SgRNA	PAM/PFS	检测平台 Detection platform	靶向目标 Target type	信号放大方法 Signal amplification	灵敏性 Sensitivity	多路复用 Multiplex	定量 Quantitative	便携性 Portable	时间 Time/h
Cas9	HNH RuvC	tracrRNA	5'NGG	无	DNA	CAS-EXPAR	aM	无	无	是	<1
Cas9	HNH RuvC	crRNA	5'NGG	无	DNA	CAS-EXPAR
Cas12a Cas12b	RuvC	crRNA	5'TTN	HOLMES	DNA	PCR; RT-PCR;RPA	aM	无	无	否	1
				DETECTR	DNA	PCR; RT-PCR;RPA	aM	无	无	是	1
				HOLMESv2	DNA	LAMP; RT-LAMP	aM	无	有	是	1
Cas13a	HEPN	crRNA	3'A、U 或C	SHERLOCK	RNA	RPA	aM	无	无	是	2
Cas14	RuvC	tracrRNA crRNA	无	DETECTR	ssDNA	RPA	aM	无	无	是	1

效应物 Effector	结构域 Domain	向导RNA SgRNA	PAM/PFS	检测平台 Detection platform	靶向目标 Target type	信号放大方法 Signal amplification	灵敏性 Sensitivity	多路复用 Multiplex	定量 Quantitative	便携性 Portable	时间 Time/h
Cas9	HNH RuvC	tracrRNA	5'NGG	无	DNA	CAS-EXPAR	aM	无	无	是	<1
Cas9	HNH RuvC	crRNA	5'NGG	无	DNA	CAS-EXPAR
Cas12a Cas12b	RuvC	crRNA	5'TTN	HOLMES	DNA	PCR; RT-PCR;RPA	aM	无	无	否	1
				DETECTR	DNA	PCR; RT-PCR;RPA	aM	无	无	是	1
				HOLMESv2	DNA	LAMP; RT-LAMP	aM	无	有	是	1
Cas13a	HEPN	crRNA	3'A、U 或C	SHERLOCK	RNA	RPA	aM	无	无	是	2
Cas14	RuvC	tracrRNA crRNA	无	DETECTR	ssDNA	RPA	aM	无	无	是	1