Application and Prospect of CRISPR/Cas System in Genetically Modified Detection

doi:10.19586/j.2095-2341.2026.0049

Current Biotechnology ›› 2026, Vol. 16 ›› Issue (4): 756-764.DOI: 10.19586/j.2095-2341.2026.0049

• Special Forum on Detection Technology for Genetically Modified Organisms • Previous Articles Next Articles

Application and Prospect of CRISPR/Cas System in Genetically Modified Detection

Jiayi ZHOU(), Lili ZHU, Gongwen HE, Dagang JIANG()

College of Life Sciences，South China Agricultural University，Guangzhou 510642，China

Received:2026-03-02 Accepted:2026-04-17 Online:2026-07-25 Published:2026-09-11
Contact: Dagang JIANG

Abstract

Abstract:

With the widespread cultivation of genetically modified crops， there is an urgent need for efficient and accurate detection technologies to protect consumer rights and enable government regulation. Although PCR-based molecular detection techniques have been widely adopted， there remains scope for improvement in on-site rapid screening， simultaneous multi-target detection， and device portability. The clustered regularly interspaced short palindromic repeats-associated protein （CRISPR/Cas） system， with its unique and powerful cleavage activity， offers a breakthrough solution for developing simple， on-site detection technologies. Toward the application of the CRISPR/Cas system in genetically modified detection， this review summarized its recent advances and current implementations. It elaborated on and analyzed the technical principles and practical applications of molecular diagnostic platforms based on Cas9， Cas12， Cas13， and Cas14 proteins， including their advantages in enhancing detection sensitivity， enabling multi-target recognition， simplifying operational procedures， and promoting device portability. The paper also summarized the current limitations of these technologies and discussed future development directions， aiming to advance the progress of rapid， precise， and portable novel detection technologies.

Key words: CRISPR/Cas, detection technology, genetically modified crops

CLC Number:

Q786

Jiayi ZHOU, Lili ZHU, Gongwen HE, Dagang JIANG. Application and Prospect of CRISPR/Cas System in Genetically Modified Detection[J]. Current Biotechnology, 2026, 16(4): 756-764.

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https://swjsjz.magtechjournal.com/EN/Y2026/V16/I4/756

Figures/Tables 4

References 49

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比较维度	PCR、RPA/LAMP-电泳	免疫层析试纸条技术	CRISPR/Cas系统技术
准确性	关键在于引物设计，存在非特异性扩增风险	关键在于抗原-抗体结合，可能导致假阴性	具有核酸引物设计与crRNA特异性识别“双重校验”机制
应用场景	需要复杂仪器，难以实现现场快速检测	可实现裸眼判读，较为便捷	可适配现场检测设备的简约化，实现不依赖热循环仪和荧光读取设备的裸眼判读
技术集成	难以实现技术集合	技术架构难以支撑与核酸扩增模块的高效集成，限制了向更高集成度的一体化系统演进	基于不同Cas蛋白特性，可构建高效、无干扰的多重检测“一锅法”系统

比较维度	PCR、RPA/LAMP-电泳	免疫层析试纸条技术	CRISPR/Cas系统技术
准确性	关键在于引物设计，存在非特异性扩增风险	关键在于抗原-抗体结合，可能导致假阴性	具有核酸引物设计与crRNA特异性识别“双重校验”机制
应用场景	需要复杂仪器，难以实现现场快速检测	可实现裸眼判读，较为便捷	可适配现场检测设备的简约化，实现不依赖热循环仪和荧光读取设备的裸眼判读
技术集成	难以实现技术集合	技术架构难以支撑与核酸扩增模块的高效集成，限制了向更高集成度的一体化系统演进	基于不同Cas蛋白特性，可构建高效、无干扰的多重检测“一锅法”系统

平台名称	效应蛋白	目标序列	扩增方式	检测限	PAM序列	检测时间	参考文献
CRISDA	Cas9	DNA	SDA	amol·L^-1水平	需要	2 h	[22]
DETECTR	Cas12a	DNA	RPA	amol·L^-1水平	TTTV	1 h内	[23]
HOLMES	Cas12a	DNA/RNA	PCR/等温扩增	amol·L^-1水平	需要	1 h	[20]
HOLMESv2	Cas12b	DNA/RNA	LAMP/不对称PCR	nmol·L^-1水平	dsDNA需要	1 h内	[24]
SHERLOCK	Cas13	RNA	RPA/RT-RPA	fmol·L^-1水平	无	30~60 min	[25]
Cas14-DETECTR	Cas14	ssDNA	RPA	amol·L^-1水平	无	-	[26]

平台名称	效应蛋白	目标序列	扩增方式	检测限	PAM序列	检测时间	参考文献
CRISDA	Cas9	DNA	SDA	amol·L^-1水平	需要	2 h	[22]
DETECTR	Cas12a	DNA	RPA	amol·L^-1水平	TTTV	1 h内	[23]
HOLMES	Cas12a	DNA/RNA	PCR/等温扩增	amol·L^-1水平	需要	1 h	[20]
HOLMESv2	Cas12b	DNA/RNA	LAMP/不对称PCR	nmol·L^-1水平	dsDNA需要	1 h内	[24]
SHERLOCK	Cas13	RNA	RPA/RT-RPA	fmol·L^-1水平	无	30~60 min	[25]
Cas14-DETECTR	Cas14	ssDNA	RPA	amol·L^-1水平	无	-	[26]

效应蛋白	核酸靶点	亚型	PAM序列要求	反式切割	参考文献
Cas9	dsDNA	Ⅱ型	严格	无	[27]
Cas12	主要为dsDNA	Ⅴ型	短TTN等序列	有	[33-35]
Cas13	RNA	Ⅵ型	低	有	[44-46]
Cas14	ssDNA	Ⅴ型	无	有	[26]

Application and Prospect of CRISPR/Cas System in Genetically Modified Detection

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