Open AccessHigh‐efficiency genome editing using a dmc1 promoter‐controlled CRISPR /Cas9 system in maize
Author(s) -
Feng Chao,
Su Handong,
Bai Han,
Wang Rui,
Liu Yalin,
Guo Xianrui,
Liu Chang,
Zhang Jing,
Yuan Jing,
Birchler James A.,
Han Fangpu
Publication year - 2018
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12920
Subject(s) - crispr , biology , cas9 , genome editing , genome , genetics , gene , promoter , computational biology , gene expression
Summary Previous studies revealed that the promoters for driving both Cas9 and sg RNA s 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 sg RNA , 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.