Targeted insertion of regulatory elements enables translational enhancement in rice

Frontiers in Plant Science

Plant Science

Chao Dong¹  Jian-Kang Zhu²  Xuesong Cao³  Yifu Tian⁴  Xinbo Li⁴  Rundong Shen⁴  Dating Zhong⁵  Qi Yao⁵  Xuening Zhang⁵  Yuming Lu⁵ 

[1] Center for Advanced Bioindustry Technologies, and Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China;Hainan Yazhou Bay Seed Lab, Sanya, Hainan, China;Center for Advanced Bioindustry Technologies, and Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China;Institute of Advanced Biotechnology, and School of Life Sciences, Southern University of Science and Technology, Shenzhen, China;Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China;Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China;Center for Advanced Bioindustry Technologies, and Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, China;Hainan Yazhou Bay Seed Lab, Sanya, Hainan, China;Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, China;Shanghai Collaborative Innovation Center of Agri-Seeds, Joint Center for Single Cell Biology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China;

关键词: translational regulation;    enhancer;    knock-in;    rice;    breeding;   

DOI  :  10.3389/fpls.2023.1134209

 received in 2022-12-30, accepted in 2023-03-13,  发布年份 2023

来源: Frontiers

【 摘 要 】

In-locus editing of agronomically-important genes to optimize their spatiotemporal expression is becoming an important breeding approach. Compared to intensive studies on mRNA transcription, manipulating protein translation by genome editing has not been well exploited. Here, we found that precise knock-in of a regulating element into the 5’UTR of a target gene could efficiently increase its protein abundance in rice. We firstly screened a translational enhancer (AMVE) from alfalfa mosaic virus using protoplast-based luciferase assays with an 8.5-folds enhancement. Then the chemically modified donor of AMVE was synthesized and targeted inserted into the 5’UTRs of two genes (WRKY71 and SKC1) using CRISPR/Cas9. Following the in-locus AMVE knock-in, we observed up to a 2.8-fold increase in the amount of WRKY71 protein. Notably, editing of SKC1, a sodium transporter, significantly increased salt tolerance in T2 seedlings, indicating the expected regulation of AMVE knock-in. These data demonstrated the feasibility of such in-locus editing to enhance protein expression, providing a new approach to manipulating protein translation for crop breeding.

【 授权许可】

Unknown   
Copyright © 2023 Shen, Yao, Zhong, Zhang, Li, Cao, Dong, Tian, Zhu and Lu