CRISPR/Cas系统在转基因作物检测中的应用和展望

doi:10.19586/j.2095-2341.2026.0049

生物技术进展 ›› 2026, Vol. 16 ›› Issue (4): 756-764.DOI: 10.19586/j.2095-2341.2026.0049

CRISPR/Cas系统在转基因作物检测中的应用和展望

周佳奕(), 朱梨梨, 何龚雯, 姜大刚()

华南农业大学生命科学学院，广州　510642

收稿日期:2026-03-02 接受日期:2026-04-17 出版日期:2026-07-25 发布日期:2026-09-11
通信作者: 姜大刚
作者简介:周佳奕 E-mail：jiayizhou@stu.scau.edu.cn；
基金资助:
农业生物育种重大专项(2023ZD04062)

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

摘要/Abstract

摘要：

随着转基因（genetically modified，GM）作物被广泛种植，为保护消费者权益和服务政府监管，亟需高效精准的转基因检测技术。基于PCR的分子检测技术应用广泛，但在现场快速筛查、多靶标同步检测及设备便携性方面仍有进一步提升的空间。成簇间隔短回文重复序列及相关蛋白（clustered regularly interspaced short palindromic repeats-associated protein，CRISPR/Cas）系统凭借其强大的切割活性，为发展现场简便的检测技术提供了突破性解决方案。为了更好地将CRISPR/Cas系统应用于转基因检测，综述了其在转基因检测中的前沿进展和应用现状，阐述并重点分析了基于Cas9、Cas12、Cas13和Cas14等蛋白的分子诊断平台的技术原理与应用实践，包括在提升检测灵敏度、实现多重靶标识别、简化操作流程及促进设备便携化方面的优势，总结了当前技术存在的不足，并对未来发展方向进行了展望，以期推动快速、精准、便携的新型检测技术的发展进程。

关键词: CRISPR/Cas, 检测技术, 转基因作物

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

中图分类号:

Q786

周佳奕, 朱梨梨, 何龚雯, 姜大刚. CRISPR/Cas系统在转基因作物检测中的应用和展望[J]. 生物技术进展, 2026, 16(4): 756-764.

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/CN/Y2026/V16/I4/756

图/表 4

参考文献 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]

CRISPR/Cas系统在转基因作物检测中的应用和展望

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

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