Targeted mutagenesis in Arabidopsis thaliana using CRISPR-Cas12b/C2c1.,Journal of Integrative Plant Biology

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Targeted mutagenesis in Arabidopsis thaliana using CRISPR-Cas12b/C2c1.
Journal of Integrative Plant Biology ( IF 9.3 ) Pub Date : 2020-05-12 , DOI: 10.1111/jipb.12944
Fan Wu ₁ , Xinyu Qiao ₁ , Yafei Zhao ₁ , Ziyi Zhang ₁ , Yifan Gao ₁ , Lingfeng Shi ₁ , Haokun Du ₁ , Lulu Wang ₁ , Ya-Jie Zhang ₂ , Yu Zhang ₃ , Langyu Liu ₂ , Quan Wang ₃ , Dejing Kong _{1,

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Affiliation

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.

中文翻译：

使用CRISPR-Cas12b / C2c1在拟南芥中进行定向诱变。

Cas12b / C2c1是新近鉴定的2类CRISPR核酸内切酶，最近被设计用于哺乳动物和水稻中的靶向基因组编辑。为了探索CRISPR‐Cas12b系统在双子叶拟南芥中的潜在应用，我们选择了BvCas12b和BhCas12b v4进行分析。我们成功地使用了两种核酸内切酶来诱导突变，执行多基因组编辑，并在多个基因座处产生大的缺失。在潜在的脱靶位点未检测到明显的突变。通过靶向基因诱变产生的突变体的插入/缺失频率和模式分析表明，CRISPR‐Cas12b系统可用于拟南芥基因组编辑。

更新日期：2020-05-12

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