Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System
Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System
复制标题
使用 TALEN 和 CRISPR/Cas 系统对玉米进行定向诱变
DOI：
10.1016/j.jgg.2013.12.001
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发表时间：
2014-02-01
影响因子：
5.9
通讯作者：
Gao, Caixia
中科院分区：
文献类型：
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作者：
Liang, Zhen;Zhang, Kang;Gao, Caixia
转录激活因子样效应核酸酶（TALENs）和聚集规律间隔短回文重复序列(CRISPR)/CRISPR相关（Cas）系统已成为多种物种基因组编辑的强大工具。在这里，我们首次报道了利用TALENs和CRISPR/Cas系统对玉米进行靶向诱变。我们设计了5个靶向ZmPDS、ZmIPK1A、ZmIPK、ZmMRP4 4个基因的TALENs，在原生质体中获得了高达23.1%的靶向效率，其中约13.3% ~ 39.1%的转基因植株为体细胞突变。此外，我们在玉米原生质体中构建了两个靶向ZmIPK基因的grna，频率分别为16.4%和19.1%。此外，CRISPR/Cas系统在Z. mays原生质体中诱导靶向突变的效率（13.1%）与TALENs获得的效率（9.1%）相似。我们的研究结果表明，TALENs和CRISPR/Cas系统都可以用于玉米基因组修饰。
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.