Application of CRISPR-Cas System in Nucleic Acid Detection

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

Abstract

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

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.

Figures/Tables 1

References 63

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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