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

作     者：Zhen Liang Kang Zhang Kunling Chen Caixia Gao 

作者机构：State Key Laboratory of Plant Cell and Chromosome EngineeringInstitute of Genetics and Developmental BiologyChinese Academy of Sciences 

出 版 物：《Journal of Genetics and Genomics》 (遗传学报（英文版）)

年 卷 期：2014年第41卷第2期

页      面：63-68页

核心收录：

学科分类：081302[工学-建筑设计及其理论] 08[工学] 09[农学] 0901[农学-作物学] 0813[工学-建筑学] 

基　　金：supported by the National Natural Science Foundation of China (Grant Nos. 31271795 and 31200273) 

主　　题：TAL-effector nucleases CRISPR/Cas system Knock-out Zea mays 

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, ZmlPKIA, ZmlPK, 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 ZmlPK 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.