Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System

被引:402
作者
Liang, Zhen [1 ]
Zhang, Kang [1 ]
Chen, Kunling [1 ]
Gao, Caixia [1 ]
机构
[1] Chinese Acad Sci, Inst Genet & Dev Biol, State Key Lab Plant Cell & Chromosome Engn, Beijing 100101, Peoples R China
基金
中国国家自然科学基金;
关键词
TAL-effector nucleases; CRISPR/Cas system; Knock-out; Zea mays; ZINC-FINGER NUCLEASES; GENOME MODIFICATION; DNA CLEAVAGE; MAIZE; CAS9; RNA; ARABIDOPSIS; GENES; SPECIFICITY; MULTIPLEX;
D O I
10.1016/j.jgg.2013.12.001
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems have emerged as powerful tools for genome editing in a variety of species. Here, we report, for the first time, targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system. We designed five TALENs targeting 4 genes, namely ZmPDS, ZmIPK1A, ZmIPK, ZmMRP4, and obtained targeting efficiencies of up to 23.1% in protoplasts, and about 13.3% to 39.1% of the transgenic plants were somatic mutations. Also, we constructed two gRNAs targeting the ZmIPK gene in maize protoplasts, at frequencies of 16.4% and 19.1%, respectively. In addition, the CRISPR/Cas system induced targeted mutations in Z. mays protoplasts with efficiencies (13.1%) similar to those obtained with TALENs (9.1%). Our results show that both TALENs and the CRISPR/Cas system can be used for genome modification in maize.
引用
收藏
页码:63 / 68
页数:6
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