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Directed evolution rice genes with randomly multiplexed sgRNAs assembly of base editors

Plant Biotechnol J. 2023-08; 
Ao Zhang, Tiaofeng Shan, Yan Sun, Zhipeng Chen, Jianjian Hu, Zhichao Hu, Ziheng Ming, Zhitao Zhu, Xue Li, Jun He, Shijia Liu, Ling Jiang, Xiaoou Dong, Yufeng Wu, Yanpeng Wang, Yuqiang Liu, Chao Li, Jianmin Wan
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Proteins, Expression, Isolation and Analysis … the primer sets used in this work are listed in Table S9 and were synthesized by GenScript. … Saturated target oligos were synthesized by GenScript. Fifty micromoles of forward and … Get A Quote

摘要

CRISPR-based directed evolution is an effective breeding biotechnology to improve agronomic traits in plants. However, its gene diversification is still limited using individual single guide RNA. We described here a multiplexed orthogonal base editor (MoBE), and a randomly multiplexed sgRNAs assembly strategy to maximize gene diversification. MoBE could induce efficiently orthogonal ABE (<36.6%), CBE (<36.0%), and A&CBE (<37.6%) on different targets, while the sgRNA assembling strategy randomized base editing events on various targets. With respective 130 and 84 targets from each strand of the 34th exon of rice acetyl-coenzyme A carboxylase (OsACC), we observed the target-scaffold combination types up to 27 2... More

关键词

directed evolution, dual base editor, herbicide resistance, multiplexed and orthogonal base editor, randomly multiplexed sgRNAs assembly
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