J Integr Plant Biol. ›› 2020, Vol. 62 ›› Issue (11): 1653-1658.DOI: 10.1111/jipb.12944

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Targeted mutagenesis in Arabidopsis thaliana using CRISPR-Cas12b/C2c1

Fan Wu1† , Xinyu Qiao1† , Yafei Zhao1† , Ziyi Zhang1 , Yifan Gao1 , Lingfeng Shi1 , Haokun Du1 , Lulu Wang1 , Ya‐Jie Zhang2 , Yu Zhang3 , Langyu Liu2 *, Quan Wang3 * and Dejing Kong1,4*   

  1. 1School of Biological Science and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, China
    2College of Life Sciences, Capital Normal University, Beijing 100048, China
    3Lingnan Laboratory of Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518120, China
    4State Key Laboratory Breeding Base‐Hebei Province Key Laboratory of Molecular Chemistry for Drug, Shijiazhuang 050018, China

    These authors contributed equally to this work.
    *Correspondences:
    Email: Langyu Liu (liangyu.liu@cnu.edu.cn); Quan Wang (wangquan03@caas.cn); Dejing Kong (kongdejing@hebust.edu.cn, Dr. Kong is fully responsible for the distribution of all materials associated with this article)
  • Received:2020-04-24 Accepted:2020-05-08 Online:2020-05-12 Published:2020-11-01

Abstract:

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.

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