RPA-CRISPR/Cas12b技术结合侧向流动试纸条检测基因编辑水稻

doi:10.3969/j.issn.1674-7968.2025.10.019

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Abstract Currently, the detection of gene editing products mostly relies on methods such as genome sequencing analysis and PCR technology, which are time-consuming, costly, and require large instruments and equipment. In order to establish a rapid, sensitive, visual and suitable method for the detection of gene edited crops, primers and single guide RNA (sgRNA) were designed according to the rice OsSWEET14 gene editing material. Recombinase polymerase amplification (RPA) and CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated protein) 12b technology combined with lateral flow strips (LFS) strip were used to detect gene edited rice (Oryza sativa). The results showed that RPA-CRISPR/Cas12b combined with LFS could make the detection results visualized through the test strips within 33 min at most, and the detection sensitivity was 100 fg/μL. The RPA-CRISPR/Cas12b-LFS detection method developed in this study was simple and rapid, did not need large instruments and equipment. This study provides technical support for the field rapid detection with limited experimental conditions.

Key words： Rapid detection Recombinase polymerase amplification Lateral flow strips CRISPR/Cas12b Gene editing

Received: 19 November 2024

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Corresponding Authors: *zhaijf@caiq.org.cn

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	Articles by authors
	HU Guang
	LUO Liang
	ZHAO Wen-Jun
	WANG Zhi
	FU Wei
	YU Yan-Xue
	TIAN Qian
	WANG Meng-Yu
	ZHAI Jun-Feng

Cite this article:

HU Guang,LUO Liang,ZHAO Wen-Jun, et al. Detection of Gene-edited Rice (Oryza sativa) by RPA-CRISPR/Cas12b Technology Combined with Lateral Flow Strips[J]. 农业生物技术学报, 2025, 33(10): 2322-2328.

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https://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2025.10.019 OR https://journal05.magtech.org.cn/Jwk_ny/EN/Y2025/V33/I10/2322

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