A barley stripe mosaic virus-based guide RNA delivery system for targeted mutagenesis in wheat and maize
A barley stripe mosaic virus-based guide RNA delivery system for targeted mutagenesis in wheat and maize
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DOI：
10.1111/mpp.12849
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发表时间：
2019-07-05
影响因子：
4.9
通讯作者：
Zhang, Yongliang
中科院分区：
文献类型：
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作者：
Hu, Jiacheng;Li, Shaoya;Zhang, Yongliang
与常规组成型启动子驱动的表达相比，基于植物RNA病毒的向导RNA（gRNA）递送具有显著的优点，这是由于在病毒复制和移动期间gRNA的快速和稳健的扩增。到目前为止，还没有为小麦和玉米开发出病毒诱导的基因组编辑工具。在这项研究中，我们设计了一种基于大麦条纹花叶病毒（BSMV）的gRNA递送系统，用于在小麦和玉米中进行成簇规则间隔短回文重复序列（CRISPR）/Cas9介导的靶向诱变。用于靶向诱变的单个gRNA的基于BSMV的递送首先在本氏烟草中验证。为了扩展这项工作，我们用Cas9核酸酶基因转化小麦和玉米，并选择小麦TaGASR 7和玉米ZmTMS 5基因作为靶标，以评估BSMV介导的诱变的可行性和效率。对小麦和玉米的TaGASR 7和ZmTMS 5基因的正向靶向诱变达到了高达78%和48%的效率。我们的研究结果为快速有效地将gRNA递送到经济上重要的作物中提供了有用的工具。
Plant RNA virus-based guide RNA (gRNA) delivery has substantial advantages compared to that of the conventional constitutive promoter-driven expression due to the rapid and robust amplification of gRNAs during virus replication and movement. To date, virus-induced genome editing tools have not been developed for wheat and maize. In this study, we engineered a barley stripe mosaic virus (BSMV)-based gRNA delivery system for clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated targeted mutagenesis in wheat and maize. BSMV-based delivery of single gRNAs for targeted mutagenesis was first validated in Nicotiana benthamiana. To extend this work, we transformed wheat and maize with the Cas9 nuclease gene and selected the wheat TaGASR7 and maize ZmTMS5 genes as targets to assess the feasibility and efficiency of BSMV-mediated mutagenesis. Positive targeted mutagenesis of the TaGASR7 and ZmTMS5 genes was achieved for wheat and maize with efficiencies of up to 78% and 48%. Our results provide a useful tool for fast and efficient delivery of gRNAs into economically important crops.