Plant Biotechnology Journal
Abstract
Previous 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 re‐sequencing 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.
| 论文编号: | DOI:10.1111/pbi.12920 |
| 论文题目: | High Efficiency Genome Editing Using a Dmc1 Promoter‐Controlled CRISPR/Cas9 System in Maize |
| 英文论文题目: | High Efficiency Genome Editing Using a Dmc1 Promoter‐Controlled CRISPR/Cas9 System in Maize |
| 第一作者: | Chao Feng, Handong Su, Bai Han, Rui Wang, Yalin Liu, Xianrui Guo, Chang Liu, Jing Zhang, Jing Yuan, James A. Birchler, Fangpu Han |
| 英文第一作者: | Chao Feng, Handong Su, Bai Han, Rui Wang, Yalin Liu, Xianrui Guo, Chang Liu, Jing Zhang, Jing Yuan, James A. Birchler, Fangpu Han |
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| 发表年度: | 2018-03-26 |
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| 摘要: | Previous 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 re‐sequencing 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. |
| 英文摘要: | Previous 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 re‐sequencing 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. |
| 刊物名称: | Plant Biotechnology Journal |
| 英文刊物名称: | Plant Biotechnology Journal |
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| 其它备注: | Chao Feng, Handong Su, Bai Han, Rui Wang, Yalin Liu, Xianrui Guo, Chang Liu, Jing Zhang, Jing Yuan, James A. Birchler, Fangpu Han. High Efficiency Genome Editing Using a Dmc1 Promoter‐Controlled CRISPR/Cas9 System in Maize. Plant Biotechnology Journal. DOI:10.1111/pbi.12920 |
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