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High-throughput genome editing in rice with a virus-based surrogate system

J Integr Plant Biol. 2022-10; 
Yifu Tian, Dating Zhong, Xinbo Li, Rundong Shen, Han Han, Yuqin Dai, Qi Yao, Xuening Zhang, Qi Deng, Xuesong Cao, Jian-Kang Zhu, Yuming Lu
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Proteins, Expression, Isolation and Analysis … Western blotting was performed with a mouse anti-FLAG antibody (1:2,000 dilution, Sangon Biotech, China) or mouse anti-SpCas9 antibody (1:1,000 dilution, Genscript, USA). The HRP… Get A Quote

摘要

With the widespread use of CRISPR/Cas technologies in plants, large-scale genome editing is increasingly needed. Here, we developed a geminivirus-mediated surrogate system, called WDV-Gate (Wheat Dwarf Virus-surrogate), to facilitate high-throughput genome editing. WDV-Gate has two parts: one is the recipient callus from a transgenic rice line expressing Cas9 and a mutated hygromycin-resistant gene (HygM) for surrogate selection; the other is a WDV-based construct expressing two sgRNAs targeting HygM and a gene of interest, respectively. We evaluated WDV-Gate on six rice loci by producing a total of 874 T plants. Compared with the conventional method, the WDV-Gate system, which was characterized by a transient ... More

關鍵詞

genome editing, high-throughput, protein tagging, rice
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