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双通道检测系统使用Pyrococcus furiosus Argonaute用于大米单核酸变异量化

Ruxin Pan^1,2, Yao Yang¹, Zhiwen Lu²

¹Key Laboratory of Agricultural Genetically Modified Organisms Traceability of the Ministry of Agriculture and Rural Affairs, Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, China.

Journal of agricultural and food chemistry

|December 10, 2025

概括

使用Pyrococcus furiosus Argonaute (PfAgo) 的新型双通道检测系统精确量化了基因组编辑大米中的单核酸变异 (SNV). 这一进步为基因编辑产品提供了敏感和可靠的可追溯性.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
农业科学农业科学

背景情况:

基因组编辑技术使米等作物能够进行精确的基因修改.
精确量化单核酸变异 (SNV) 对于评估基因组编辑产品至关重要.
现有的检测方法可能缺乏复杂基因组分析所需的灵敏度或特异性.

研究的目的:

开发一种敏感和特定的双通道检测系统,用于对基因组编辑大米的SNV量化.
利用Pyrococcus furiosus Argonaute (PfAgo) 来同时检测野生类型和SNV目标.
建立一个可靠的分析平台,以控制转基因大米的质量和可追溯性.

主要方法:

开发一种使用Pyrococcus furiosus Argonaute (PfAgo) 的双通道检测系统.
引入通用指导DNA (gDNA) 以同时识别野生类型和SNV目标.
设计具有特定不匹配的分子信标,以防止交叉干扰并实现精确量化.

主要成果:

该系统实现了SNV突变的0.1%的敏感检测极限.
在各种基因组编辑的米变种中展示了单核酸歧视能力.
在盲测试中表现出强大的定量可靠性,相对标准偏差低 (<6%) 和偏差低 (<±8%).

结论:

开发的基于PfAgo的双通道系统提供了高灵敏度,特异性和SNVs的精确量化.
这种方法为基因组编辑大米的可追溯性和质量控制提供了强大的分析工具.
简化的系统设计提高了在作物改进计划中检测遗传变异的效率.

关键词:

这是一种Pyrococcus furiosus argonaute.采用双通道检测系统进行检测.基因组编辑的生物体量化量化量化量化量化量化量化量化量化量化量化量单核酸变异是一种单核酸变异

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