Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize

作者：Yuanyuan Jiang, Yiping Chai, Minhui Lu, Xiuli Han, Qiupeng Lin, Yingxin Zhang, Qiang Zhang, Yun Zhou, Xuechen Wang, Caixia Gao, Qijun Chen · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2020 · DOI：10.1101/2020.07.06.188896 · 被引用次数：34 · 研究领域：CRISPR and Genetic Engineering、Chromosomal and Genetic Variations、Plant Virus Research Studies

Abstract A novel and universal CRISPR/Cas-derived precision genome-editing technology named prime editing was developed. However, low efficiency of prime editing was shown in transgenic rice lines. We reasoned that enhancing pegRNA expression would improve prime-editing efficiency. In this report, we describe two strategies for enhancing pegRNA expression. We constructed 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 achieved much higher prime-editing efficiency in maize. Our results are inspiring and provide a direction for the optimization of plant prime editors.