Lou Huanhuan, Xiang Haiying, Zeng Wanli, Jiang Jiarui, Zhang Jianduo, Xu Li, Zhao Chenglu, Gao Qian, Li Zhenghe
Institute of Biotechnology, Zhejiang University, Hangzhou, Zhejiang 310058, China.
Yunnan Academy of Tobacco Science, Kunming, Yunnan 650106, China.
STAR Protoc. 2024 Dec 20;5(4):103437. doi: 10.1016/j.xpro.2024.103437. Epub 2024 Nov 5.
Plant virus vectors have emerged as promising tools for CRISPR-Cas reagent delivery. Here, we present a protocol for DNA-free plant genome editing using an engineered RNA virus vector for the transient delivery of CRISPR-Cas components. We describe steps for viral vector construction, viral vector recovery through agroinoculation of Nicotiana benthamiana, mechanical inoculation of target plant hosts, analysis of somatic mutagenesis frequency, and regeneration of mutant plants. The method achieves high editing efficiency and eliminates the need for stable plant transformation. For complete details on the use and execution of this protocol, please refer to Liu et al..
植物病毒载体已成为用于递送CRISPR-Cas试剂的有前景的工具。在此，我们展示了一种使用工程化RNA病毒载体进行无DNA植物基因组编辑的方案，用于CRISPR-Cas组件的瞬时递送。我们描述了病毒载体构建、通过农杆菌接种本氏烟草回收病毒载体、对目标植物宿主进行机械接种、分析体细胞诱变频率以及突变植物再生的步骤。该方法实现了高编辑效率，并且无需进行稳定的植物转化。有关本方案使用和实施的完整详细信息，请参考Liu等人的文献。
