High‐efficiency genome editing using a dmc1 promoter‐controlled CRISPR/Cas9 system in maize

• DOI: 10.1111/pbi.12920
• Publication Date: 2018-03-23T11:28:54Z
• Authors: Chao Feng; Handong Su; Han Bai; Rui Wang; Yalin Liu; Xianrui Guo; Chang Liu; Jing Zhang; Jing Yuan; James A. Birchler; Fangpu Han
• Keywords: Gene Editing; 0301 basic medicine; 2. Zero hunger; Genome; targeted genome editing; Genetically Modified; Plant; Plants; maize; Plants, Genetically Modified; Zea mays; Promoter Regions; 03 medical and health sciences; Genetic; dmc1 promoter; CRISPR-Associated Protein 9; Mutation; CRISPR-Cas Systems; Promoter Regions, Genetic; CRISPR/Cas9; Research Articles; Genome, Plant

SummaryPrevious studies revealed that the promoters for driving both Cas9 and sgRNAs are quite important for efficient genome editing by CRISPR/Cas9 in plants. Here, we report our results of targeted genome editing using the maize dmc1 gene promoter combined with the U3 promoter for Cas9 and sgRNA, respectively. Three loci in the maize genome were selected for targeting. The T0 plants regenerated were highly efficiently edited at the target sites with homozygous or bi‐allelic mutants accounting for about 66%. The mutations in T0 plants could be stably transmitted to the T1 generation, and new mutations could be generated in gametes or zygotes. Whole‐genome resequencing indicated that no off‐target mutations could be detected in the predicted loci with sequence similarity to the targeted site. Our results show that the dmc1 promoter‐controlled (DPC) CRISPR/Cas9 system is highly efficient in maize and provide further evidence that the optimization of the promoters used for the CRISPR/Cas9 system is important for enhancing the efficiency of targeted genome editing in plants. The evolutionary conservation of the dmc1 gene suggests its potential for use in other plant species.