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Generation of targeted mutant rice using a CRISPR-Cpf1 system

文献类型： 外文期刊

作者： Xu, Rongfang 1 ; Qin, Ruiying 1 ; Li, Hao 1 ; Li, Dongdong 1 ; Li, Li 1 ; Wei, Pengcheng 1 ; Yang, Jianbo 1 ;

作者机构： 1.Anhui Acad Agr Sci, Rice Res Inst, Key Lab Rice Genet Breeding Anhui Prov, Hefei, Peoples R China

关键词： genome editing;Cpf1;CRISPR;rice

期刊名称：PLANT BIOTECHNOLOGY JOURNAL （ 影响因子：9.803； 五年影响因子：9.555 ）

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收录情况： SCI

摘要： CRISPR-Cpf1 is a newly identified CRISPR-Cas system, and Cpf1 was recently engineered as a molecular tool for targeted genome editing in mammalian cells. To test whether the engineered CRISPR-Cpf1 system could induce the production of rice mutants, we selected two genome targets in the OsPDS and OsBEL genes. Our results show that both targets could be efficiently mutated in transgenic rice plants using CRISPR-Cpf1. We found that pre-crRNAs with a full-length direct repeat sequence exhibited considerably increased efficiencies compared with mature crRNAs. In addition, the specificity and transmission of the mutation were investigated, and the behaviours of crRNA-Cpf1-induced plant targeted genome mutagenesis were assessed. Taken together, our results indicate that CRISPR-Cpf1 expression via stable transformation can efficiently generate specific and heritable targeted mutations in rice and thereby constitutes a novel and important approach to specific and precise plant genome editing.

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