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Prime Editing Efficiently Generates W542L and S621I Double Mutations in Two ALS Genes in Maize

Yuanyuan Jiang, Yiping Chai, Minhui Lu, Xiuli Han, Qiupeng Lin, Yu Zhang, Qiang Zhang, Yun Zhou, Xuechen Wang, Caixia Gao & Qijun Chen


Genome Biology
DOI:10.1186/s13059-020-02170-5


Abstract

Prime editing is a novel and universal CRISPR/Cas-derived precision genome-editing technology that has been recently developed. However, low efficiency of prime editing has been shown in transgenic rice lines. We hypothesize that enhancing pegRNA expression could improve prime-editing efficiency. In this report, we describe two strategies for enhancing pegRNA expression. We construct a prime editing vector harboring two pegRNA variants for W542L and S621I double mutations in ZmALS1 and ZmALS2. Compared with previous reports in rice, we achieve much higher prime-editing efficiency in maize. Our results are inspiring and provide a direction for the optimization of plant prime editors.
 
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