Journal of Genetics and Genomics
Abstract
MAD7 is an engineerednucleaseof the Class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) family with a low level of homology to canonical Cas12anucleases. It has been publicly released as a royalty-free nuclease for both academic and commercial use. Here, we demonstrate that the CRISPR-MAD7 system can be used for genome editingand recognizes T-rich PAM sequences (YTTN) in plants. Its editing efficiency in rice and wheat is comparable to that of the widely used CRISPR-LbCas12a system. We developed two variants, MAD7-RR and MAD7-RVR, that increase the target range of MAD7, as well as an M-AFID (a MAD7-APOBEC fusion-induced deletion) system that creates predictable deletions from 5′-deaminated Cs to the MAD7-cleavage site. Moreover, we show that MAD7 can be used for multiplex gene editing and that it is effective in generatingindelswhen combined with otherCRISPRRNA orthologs. Using the CRISPR-MAD7 system, we have obtained regenerated mutant rice and wheat plants with up to 65.6% efficiency.
| 论文编号: | DOI:10.1016/j.jgg.2021.04.003 |
| 论文题目: | Genome Editing in Plants with MAD7 Nuclease |
| 英文论文题目: | Genome Editing in Plants with MAD7 Nuclease |
| 第一作者: | Qiupeng Lin, Zixu Zhu, Guanwen Liu, Chao Sun, Dexing Lin, Chenxiao Xue, Shengnan Li, Dandan Zhang, Caixia Gao, Yanpeng Wang, Jin-Long Qiu |
| 英文第一作者: | Qiupeng Lin, Zixu Zhu, Guanwen Liu, Chao Sun, Dexing Lin, Chenxiao Xue, Shengnan Li, Dandan Zhang, Caixia Gao, Yanpeng Wang, Jin-Long Qiu |
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| 发表年度: | 2021-07-08 |
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| 摘要: | MAD7 is an engineerednucleaseof the Class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) family with a low level of homology to canonical Cas12anucleases. It has been publicly released as a royalty-free nuclease for both academic and commercial use. Here, we demonstrate that the CRISPR-MAD7 system can be used for genome editingand recognizes T-rich PAM sequences (YTTN) in plants. Its editing efficiency in rice and wheat is comparable to that of the widely used CRISPR-LbCas12a system. We developed two variants, MAD7-RR and MAD7-RVR, that increase the target range of MAD7, as well as an M-AFID (a MAD7-APOBEC fusion-induced deletion) system that creates predictable deletions from 5′-deaminated Cs to the MAD7-cleavage site. Moreover, we show that MAD7 can be used for multiplex gene editing and that it is effective in generatingindelswhen combined with otherCRISPRRNA orthologs. Using the CRISPR-MAD7 system, we have obtained regenerated mutant rice and wheat plants with up to 65.6% efficiency. |
| 英文摘要: | MAD7 is an engineerednucleaseof the Class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) family with a low level of homology to canonical Cas12anucleases. It has been publicly released as a royalty-free nuclease for both academic and commercial use. Here, we demonstrate that the CRISPR-MAD7 system can be used for genome editingand recognizes T-rich PAM sequences (YTTN) in plants. Its editing efficiency in rice and wheat is comparable to that of the widely used CRISPR-LbCas12a system. We developed two variants, MAD7-RR and MAD7-RVR, that increase the target range of MAD7, as well as an M-AFID (a MAD7-APOBEC fusion-induced deletion) system that creates predictable deletions from 5′-deaminated Cs to the MAD7-cleavage site. Moreover, we show that MAD7 can be used for multiplex gene editing and that it is effective in generatingindelswhen combined with otherCRISPRRNA orthologs. Using the CRISPR-MAD7 system, we have obtained regenerated mutant rice and wheat plants with up to 65.6% efficiency. |
| 刊物名称: | Journal of Genetics and Genomics |
| 英文刊物名称: | Journal of Genetics and Genomics |
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| 其它备注: | Qiupeng Lin, Zixu Zhu, Guanwen Liu, Chao Sun, Dexing Lin, Chenxiao Xue, Shengnan Li, Dandan Zhang, Caixia Gao, Yanpeng Wang, Jin-Long Qiu. Genome Editing in Plants with MAD7 Nuclease. Journal of Genetics and Genomics. DOI:10.1016/j.jgg.2021.04.003 |
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