Targeted mutagenesis in Arabidopsis thaliana using CRISPR‐Cas12b/C2c1

作者：Fan Wu, Xinyu Qiao, Yafei Zhao, Ziyi Zhang, Yifan Gao, Lingfeng Shi, Haokun Du, Lulu Wang, Yajie Zhang, Yu Zhang, Langyu Liu, Quan Wang, Dejing Kong · 发表于：Journal of Integrative Plant Biology · 年份：2020 · DOI：10.1111/jipb.12944 · 被引用次数：44 · 研究领域：CRISPR and Genetic Engineering、Insect symbiosis and bacterial influences、Plant Virus Research Studies

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.