中国农业科学2024，Vol.57Issue(17)：3318-3334,17.DOI:10.3864/j.issn.0578-1752.2024.17.002
农业用植物基因编辑产品检测方法研究进展
Progress on Detection Methods for Gene-Edited Organisms
摘要
Abstract
Gene editing techniques have made gene edited(GE)organisms enter commercial applications from laboratories.In 2022,the Ministry of Agriculture and Rural Affairs specifically issued the"Guidelines for Safety Evaluation of Genetically Edited Plants for Agricultural Use(Trial)"for the safety evaluation of GE plants without introducing exogenous genes.In 2023,China granted the first biosafety certificate for GE soybean AE15-18-1,marking the official start of the commercialization process of GE crops in China.GE organisms are different from traditional genetically modified organisms(GMOs)containing exogenous DNA sequences,making common GM detection strategies inapplicable to the detection of GE organisms.As the industrialization of GE crops progresses positively,how to efficiently and accurately detect whether a product is gene-edited and its editing characteristics is an important basis for the commercial use and intellectual property protection of GE products.There is an urgent need to develop detection technologies suitable for GE products.With the goal of detecting whether the target sequence has been edited,many detection technologies have been developed based on PCR,sequencing,and other technologies,and are widely used in the screening of GE products in the research and development process.After industrialization,safety supervision and intellectual property protection require not only the detection of whether the sample has been edited but also the rapid identification of the nucleotide sequence characteristics of the sample to determine its origin and identity.Subsequently,precise quantification of the GE components is necessary to determine whether quantitative labeling is required.Currently,it is difficult to quickly identify the identity of GE products with only a few base insertions,deletions,and single nucleotide variations(SNV)using conventional PCR or sequencing technologies.It is even more challenging to accurately quantify the content of GE components.Aiming at the rapid identification of the DNA sequence characteristics after editing and precise quantification,based on the molecular characteristics of GE products,this paper reviews the application of the gel electrophoresis-based PCR method,the sequencing-based method,the real-time PCR-based method,the digital PCR-based method,the editing enzyme-based method,and the instrument-based method in detection of GE organisms,and expounds the advantages and disadvantages of each method during detection.This review initially explores the detection and quantification strategies suitable for GE organisms and provides a reference for subsequent development of detection methods for GE organisms.关键词
基因编辑/测序/实时荧光PCR/数字PCR/可编程内切酶Key words
gene editing/sequencing/real time PCR/digital PCR/editing enzyme引用本文复制引用
武玉花,徐利群,翟杉杉,普豪祯,高鸿飞,张华,李俊,李允静,肖芳,吴刚..农业用植物基因编辑产品检测方法研究进展[J].中国农业科学,2024,57(17):3318-3334,17.基金项目
科技创新2030—重大项目(2022ZD0402010) （2022ZD0402010）
