基于TaqMan-qPCR技术的OsWx基因编辑水稻检测方法的建立  

Establishment of Detection Method Based on TaqMan Real-time Fluorescence Quantitative PCR Technology for OsWx-edited Rice

作　　者：胡广 王智 付伟 石雨婷 陈珊珊 罗亮 魏霜 HU Guang;WANG Zhi;FU Wei;SHI Yuting;CHEN Shanshan;LUO Liang;WEI Shuang(Technology Center of Guangzhou Customs District,Guangzhou 510423,China;Chinese Academy of Inspection and Quarantine,Beijing 100176,China;Development Center of Science and Technology,Ministry of Agriculture and Rural Affairs,Beijing 100176,China;College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]广州海关技术中心,广州510423 [2]中国检验检疫科学研究院,北京100176 [3]农业农村部科技发展中心,北京100176 [4]湖南农业大学生物科学技术学院,长沙410128

出　　处：《生物技术进展》2025年第1期86-92,共7页Current Biotechnology

基　　金：广东省基础与应用基础研究基金项目(2022A1515110843)。

摘　　要：基因编辑技术已成为当前一个重要的分子工具,通过定向改造基因组序列,在基因功能分析和作物遗传育种等方面具有广泛应用。随着基因编辑技术的快速发展,当前其可对受体生物实现几乎无痕编辑,这增加了对基因编辑产品的检测难度。因此,建立基因编辑产品的检测方法,有利于加强对市场上基因编辑产品的监管。以OsWx基因CRISPR/Cas9基因编辑材料为研究对象,设计编辑位点的特异性引物和探针进行TaqMan实时荧光定量PCR(TaqMan-qPCR)检测。研究所建立的检测体系具有良好的特异性和灵敏度,能够有效地区分OsWx单碱基突变体与野生型水稻。Gene editing technology currently has become an important molecular tool,which can selectively modify genome se⁃quences and has wide applications in gene function analysis and crop genetic breeding.Due to the rapid development of gene edit⁃ing technology,gene editing techniques can perform almost seamless editing on receptor organisms,which increases the difficul⁃ty of detecting the gene editing product.Therefore,the establishment of detection methods for gene editing products is conducive to strengthening market supervision of the gene editing product.This study was based on TaqMan real-time fluorescence quantita⁃tive PCR(TaqMan-qPCR)technology for detection to design specific primers and probes for the editing site of OsWx gene by de⁃veloped with CRISPR/Cas9.In this study,the detection system had good specificity and sensitivity,and could effectively distin⁃guish OsWx single base mutants from wild-type rice.

关 键 词：基因编辑 TaqMan-qPCR CRISPR/Cas9 OsWx 

分 类 号：Q812[生物学—生物工程] S300[农业科学—农艺学]