利用 TALENs 和 CRISPR/Cas 系统在玉米中进行靶向诱变。
Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system.
机构信息
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
出版信息
J Genet Genomics. 2014 Feb 20;41(2):63-8. doi: 10.1016/j.jgg.2013.12.001. Epub 2013 Dec 14.
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.
转录激活因子样效应物核酸酶（TALENs）和成簇规律间隔短回文重复序列（CRISPR）/CRISPR 相关（Cas）系统已成为在多种物种中进行基因组编辑的强大工具。在这里，我们首次报道了使用 TALENs 和 CRISPR/Cas 系统在玉米中进行的靶向诱变。我们设计了 5 个针对 4 个基因的 TALENs，即 ZmPDS、ZmIPK1A、ZmIPK 和 ZmMRP4，在原生质体中的靶向效率高达 23.1%，约 13.3%至 39.1%的转基因植物为体细胞突变。此外，我们还在玉米原生质体中构建了两个靶向 ZmIPK 基因的 gRNA，效率分别为 16.4%和 19.1%。此外，CRISPR/Cas 系统在玉米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。我们的结果表明，TALENs 和 CRISPR/Cas 系统均可用于玉米的基因组修饰。
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