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Targeted mutagenesis in Arabidopsis thaliana using CRISPR‐Cas12b/C2c1
Author(s) -
Wu Fan,
Qiao Xinyu,
Zhao Yafei,
Zhang Ziyi,
Gao Yifan,
Shi Lingfeng,
Du Haokun,
Wang Lulu,
Zhang YaJie,
Zhang Yu,
Liu Langyu,
Wang Quan,
Kong Dejing
Publication year - 2020
Publication title -
journal of integrative plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.734
H-Index - 83
eISSN - 1744-7909
pISSN - 1672-9072
DOI - 10.1111/jipb.12944
Subject(s) - crispr , genome editing , arabidopsis , mutagenesis , arabidopsis thaliana , biology , genetics , genome , gene , computational biology , multiplex , mutant , insertional mutagenesis , endonuclease , tilling , cas9
Cas12b/C2c1 is a newly identified class 2 CRISPR endonuclease that was recently engineered for targeted genome editing in mammals and rice. To explore the potential applications of the CRISPR‐Cas12b system in the dicot Arabidopsis thaliana , we selected BvCas12b and BhCas12b v4 for analysis. We successfully used both endonucleases to induce mutations, perform multiplex genome editing, and create large deletions at multiple loci. No significant mutations were detected at potential off‐target sites. Analysis of the insertion/deletion frequencies and patterns of mutants generated via targeted gene mutagenesis highlighted the potential utility of CRISPR‐Cas12b systems for genome editing in Arabidopsis .