文章基本信息

标题：An Editing-Site-Specific PCR Method for Detection and Quantification of CAO1-Edited Rice
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作者：Hongwen Zhang ; Jun Li ; Shengbo Zhao 等
期刊名称：Foods
电子版ISSN：2304-8158
出版年度：2021
卷号：10
期号：6
页码：1209
DOI：10.3390/foods10061209
出版社：MDPI Publishing
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.
关键词：genome-edited plants; CAO1 -edited rice; editing-site-specific PCR; identification; quantification genome-edited plants ; CAO1 -edited rice ; editing-site-specific PCR ; identification ; quantification