Genome editing in plants with MAD7 nuclease  

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 

机构地区：[1]State Key Laboratory of Plant Cell and Chromosome Engineering,Center for Genome Editing,Institute of Genetics and Developmental Biology,Innovation Academy for Seed Design,Chinese Academy of Sciences,Beijing 100101,China [2]College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing 100049,China [3]State Key Laboratory of Plant Genomics,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China [4]CAS Center for Excellence in Biotic Interactions,University of Chinese Academy of Sciences,Beijing 100049,China

出　　处：《Journal of Genetics and Genomics》2021年第6期444-451,共8页遗传学报（英文版）

基　　金：supported by grants from the Strategic Priority Research Program of the Chinese Academy of Sciences(Precision Seed Design and Breeding,XDA24020101 and XDA24020310);the National Natural Science Foundation of China(31672015,31788103);the Youth Innovation Promotion Association of the Chinese Academy of Sciences(2020000003)。

摘　　要：MAD7 is an engineered nuclease of the Class 2 type V-A CRISPR-Cas(Cas12 a/Cpf1)family with a low level of homology to canonical Cas12 a 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-Lb Cas12 a system.We develop 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 50-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.

关 键 词：MAD7 nuclease CRISPR-Cas12a Plant genome editing Royalty-free Commercial use 

分 类 号：Q943.2[生物学—植物学]