Targeted mutagenesis inArabidopsis thalianausing CRISPR-Cas12b/C2c1
Targeted mutagenesis inArabidopsis thalianausing CRISPR-Cas12b/C2c1
复制标题
使用 CRISPR-Cas12b/C2c1 对拟南芥进行靶向诱变
DOI：
10.1111/jipb.12944
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发表时间：
2020-06-19
影响因子：
11.4
通讯作者：
Kong, Dejing
中科院分区：
文献类型：
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作者：
Wu, Fan;Qiao, Xinyu;Kong, Dejing
Cas12b/C2c1是新近发现的一种新发现的2类CRISPR内切酶，最近被设计用于哺乳动物和水稻的靶向基因组编辑。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。我们成功地使用了这两种内切酶来诱导突变，执行多重基因组编辑，并在多个座位上产生大的缺失。在潜在的非靶点位置没有检测到显著的突变。通过对靶基因突变产生的突变体的插入/缺失频率和模式的分析，突出了CRISPR-Cas12b系统在拟南芥基因组编辑中的潜在用途。
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 dicotArabidopsis 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 inArabidopsis.