Open AccessGeneration of targeted mutant rice using a CRISPR ‐Cpf1 system
Author(s) -
Xu Rongfang,
Qin Ruiying,
Li Hao,
Li Dongdong,
Li Li,
Wei Pengcheng,
Yang Jianbo
Publication year - 2017
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.12669
Subject(s) - crispr , biology , genome editing , mutagenesis , genome , mutant , gene , genetics , computational biology , rna
Summary 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 Os PDS and Os BEL genes. Our results show that both targets could be efficiently mutated in transgenic rice plants using CRISPR ‐Cpf1. We found that pre‐cr RNA s with a full‐length direct repeat sequence exhibited considerably increased efficiencies compared with mature cr RNA s. In addition, the specificity and transmission of the mutation were investigated, and the behaviours of cr RNA ‐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.