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Precise, Predictable Multi-nucleotide Deletions in Rice and Wheat Using APOBEC–Cas9
Shengxing Wang, Yuan Zong, Qiupeng Lin, Huawei Zhang, Zhuangzhuang Chai, Dandan Zhang, Kunling Chen, Jinlong Qiu and Caixia Gao
Nature Biotechnology
Abstract
Short insertions and deletions can be produced in plant genomes using CRISPR–Cas editors, but reliable production of larger deletions in specific target sites has proven difficult to achieve. We report the development of a series of APOBEC–Cas9 fusion-induced deletion systems (AFIDs) that combine Cas9 with human APOBEC3A (A3A), uracil DNA-glucosidase and apurinic or apyrimidinic site lyase. In rice and wheat, AFID-3 generated deletions from 5′-deaminated Cs to the Cas9-cleavage site. Approximately one-third of deletions produced using AFID-3 in rice and wheat protoplasts (30.2%) and regenerated plants (34.8%) were predictable. We show that eAFID-3, in which the A3A in AFID-3 is replaced with truncated APOBEC3B (A3Bctd), produced more uniform deletions from the preferred TC motif to the double-strand break. AFIDs could be applied to study regulatory regions and protein domains to improve crop plants.
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DOI:10.1038/s41587-020-0566-4 |
论文题目: |
Precise, Predictable Multi-nucleotide Deletions in Rice and Wheat Using APOBEC–Cas9 |
英文论文题目: |
Precise, Predictable Multi-nucleotide Deletions in Rice and Wheat Using APOBEC–Cas9 |
第一作者: |
Shengxing Wang, Yuan Zong, Qiupeng Lin, Huawei Zhang, Zhuangzhuang Chai, Dandan Zhang, Kunling Chen, Jinlong Qiu and Caixia Gao |
英文第一作者: |
Shengxing Wang, Yuan Zong, Qiupeng Lin, Huawei Zhang, Zhuangzhuang Chai, Dandan Zhang, Kunling Chen, Jinlong Qiu and Caixia Gao |
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2020-06-30 |
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Short insertions and deletions can be produced in plant genomes using CRISPR–Cas editors, but reliable production of larger deletions in specific target sites has proven difficult to achieve. We report the development of a series of APOBEC–Cas9 fusion-induced deletion systems (AFIDs) that combine Cas9 with human APOBEC3A (A3A), uracil DNA-glucosidase and apurinic or apyrimidinic site lyase. In rice and wheat, AFID-3 generated deletions from 5′-deaminated Cs to the Cas9-cleavage site. Approximately one-third of deletions produced using AFID-3 in rice and wheat protoplasts (30.2%) and regenerated plants (34.8%) were predictable. We show that eAFID-3, in which the A3A in AFID-3 is replaced with truncated APOBEC3B (A3Bctd), produced more uniform deletions from the preferred TC motif to the double-strand break. AFIDs could be applied to study regulatory regions and protein domains to improve crop plants. |
英文摘要: |
Short insertions and deletions can be produced in plant genomes using CRISPR–Cas editors, but reliable production of larger deletions in specific target sites has proven difficult to achieve. We report the development of a series of APOBEC–Cas9 fusion-induced deletion systems (AFIDs) that combine Cas9 with human APOBEC3A (A3A), uracil DNA-glucosidase and apurinic or apyrimidinic site lyase. In rice and wheat, AFID-3 generated deletions from 5′-deaminated Cs to the Cas9-cleavage site. Approximately one-third of deletions produced using AFID-3 in rice and wheat protoplasts (30.2%) and regenerated plants (34.8%) were predictable. We show that eAFID-3, in which the A3A in AFID-3 is replaced with truncated APOBEC3B (A3Bctd), produced more uniform deletions from the preferred TC motif to the double-strand break. AFIDs could be applied to study regulatory regions and protein domains to improve crop plants. |
刊物名称: |
Nature Biotechnology |
英文刊物名称: |
Nature Biotechnology |
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Shengxing Wang, Yuan Zong, Qiupeng Lin, Huawei Zhang, Zhuangzhuang Chai, Dandan Zhang, Kunling Chen, Jinlong Qiu and Caixia Gao. Precise, Predictable Multi-nucleotide Deletions in Rice and Wheat Using APOBEC–Cas9. Nature Biotechnology. DOI:10.1038/s41587-020-0566-4 |
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