|
Genome Editing in Plants with MAD7 Nuclease
Qiupeng Lin, Zixu Zhu, Guanwen Liu, Chao Sun, Dexing Lin, Chenxiao Xue, Shengnan Li, Dandan Zhang, Caixia Gao, Yanpeng Wang, Jin-Long Qiu
Journal of Genetics and Genomics
Abstract
MAD7 is an engineered nuclease of the Class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) family with a low level of homology to canonical Cas12a nucleases. 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 editing and 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 generating indels when combined with other CRISPR RNA 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 |
联系作者: |
|
英文联系作者: |
|
外单位作者单位: |
|
英文外单位作者单位: |
|
发表年度: |
2021-07-08 |
卷: |
|
期: |
|
页码: |
|
摘要: |
MAD7 is an engineered nuclease of the Class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) family with a low level of homology to canonical Cas12a nucleases. 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 editing and 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 generating indels when combined with other CRISPR RNA 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 engineered nuclease of the Class 2 type V-A CRISPR-Cas (Cas12a/Cpf1) family with a low level of homology to canonical Cas12a nucleases. 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 editing and 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 generating indels when combined with other CRISPR RNA 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 |
论文全文: |
|
英文论文全文: |
|
全文链接: |
|
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 |
英文其它备注: |
|
学科: |
|
英文学科: |
|
影响因子: |
|
第一作者所在部门: |
|
英文第一作者所在部门: |
|
论文出处: |
|
英文论文出处: |
|
论文类别: |
|
英文论文类别: |
|
参与作者: |
|
英文参与作者: |
|
|