摘要
【目的】开发一种RUBY辅助的可视化筛选转基因植株并有效指示基因编辑发生的CRISPR/Cas9基因编辑载体GCR。【方法】将RUBY与Cas9/g RNA的表达盒偶联到一起得到GCR载体,使用水稻的OsAGO2、OsAGO3、OsAGO7为靶基因,分别构建多基因编辑载体GCR-237-1和多基因编辑载体GCR-237-2,以水稻ZH11(Zhonghua 11)为受体材料进行农杆菌介导水稻遗传转化获得基因编辑植株。【结果】GCR-237-1和GCR-237-2转化入水稻愈伤后,RUBY标记可以有效指示转基因阳性事件,并且稳定转化得到的红色植株的靶基因被高效编辑。【结论】利用基因编辑载体GCR可有效编辑水稻基因且通过肉眼直观观察即可有效指示转基因乃至基因编辑植株。
【Objective】To develop a RUBY-assisted CRISPR/Cas9 gene-editing vector(GCR)enabling visual screening of transgenic plants and gene-editing events.【Method】The GCR vector was constructed by coupling RUBY with the Cas9/gRNA expression cassette.Using OsAGO2,OsAGO3,and OsAGO7 in rice(Oryza sativa)as target genes,two multi-gene editing vectors(GCR-237-1 and GCR-237-2)were designed.These vectors were introduced into rice(Zhonghua 11,ZH11)via Agrobacterium-mediated genetic transformation to generate geneedited plants.【Result】After the transformation of GCR-237-1 and GCR-237-2 into rice callus,RUBY markers effectively indicated transgenic positive events,and the target genes of stable transformed red plants were efficiently edited.【Conclusion】The GCR vector enables effective gene editing in rice,and transgenic or gene-edited plants can be rapidly identified through direct visual observation.
作者
邓美壁
严浪
詹志田
朱敏
和玉兵
DENG Mei-bi;YAN Lang;ZHAN Zhi-tian;ZHU Min;HE Yu-bing(College of Tropical Agriculture and Forestry,Hainan University,Haikou 570100;National Nanfan Research Institute(Sanya),Research Center of Breeding in Hainan,Key Laboratory of Gene Editing Technologies(Hainan),Ministry of Agriculture and Rural Affairs,Chinese Acad-emy of Agricultural Sciences(CAAS),Sanya 570024;State Key Laboratory of Crop Gene Resources and Breeding,Institute of Crop Sciences(ICS),Chinese Academy of Agricultural Sciences(CAAS),Beijing 100081;State Key Laboratory of Crop Genetics&Germplasm Enhancement and Utilization,Nanjing Agricultural University,Nanjing 210095;Ningbo Weimeng Seed Industry Co.,Ltd.,Ningbo 315100;National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070)
出处
《生物技术通报》
北大核心
2025年第8期65-73,共9页
Biotechnology Bulletin
基金
国家自然科学基金项目(32200335)
三亚中国农业科学院国家南繁研究院南繁专项(YBXM2446,YBXM2505)。
