An Editing-Site-Specific PCR Method for Detection and Quantification of CAO1-Edited Rice

文献类型： 外文期刊

第一作者： Zhang, Hongwen

作者： Zhang, Hongwen;Li, Jun;Zhao, Shengbo;Yan, Xiaohong;Si, Nengwu;Gao, Hongfei;Li, Yunjing;Zhai, Shanshan;Xiao, Fang;Wu, Gang;Wu, Yuhua

作者机构：

关键词： genome-edited plants; CAO1-edited rice; editing-site-specific PCR; identification; quantification

期刊名称：FOODS （ 影响因子：4.35； 五年影响因子：4.957 ）

ISSN：

年卷期： 2021 年 10 卷 6 期

页码：

收录情况： SCI

摘要： Genome-edited plants created by genome editing technology have been approved for commercialization. Due to molecular characteristics that differ from classic genetically modified organisms (GMOs), establishing regulation-compliant analytical methods for identification and quantification of genome-edited plants has always been regarded as a challenging task. An editing-site-specific PCR method was developed based on the unique edited sequence in CAO1-edited rice plants. Test results of seven primer/probe sets indicated that this method can identify specific CAO1-edited rice from other CAO1-edited rice and wild types of rice with high specificity and sensitivity. The use of LNA (locked nucleic acid) in a probe can efficiently increase the specificity of the editing-site-specific PCR method at increased annealing temperature which can eliminate non-specific amplification of the non-target. The genome-edited ingredient content in blinded samples at the level of 0.1% to 5.0% was accurately quantified by this method on the ddPCR platform with RSD of <15% and bias in the range of +/- 17%, meeting the performance requirements for GMO detection method. The developed editing-site-specific PCR method presents a promising detection and quantification technique for genome-edited plants with known edited sequence.

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