biostudies-literatureJiang YYNational Transgenic Research Project257https://www.ebi.ac.uk/biostudies/studies/S-EPMC7541250biostudies-literatureUnknown21(1)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.Genome biologyPrime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize.PMC75412502019ZX08010003-001-011 and 2016ZX08009002Chen QJLin QLu MHZhang YHan XLWang XCChai YPZhou YJiang YYZhang QGao CfalsePrime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize.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.2020-01-01T00:00:00Z2020 Oct2025-04-04T22:04:35.658Z2020-10-29T09:09:09ZS-EPMC75412503302363910.1186/s13059-020-02170-5