Cas13a-based multiplex RNA targeting for potato virus Y

文献类型： 外文期刊

第一作者： Zhan, Xiaohui

作者： Zhan, Xiaohui;Song, Wenlei;Li, Shengchun;Zhang, Jiang;Zhang, Fengjuan;Tu, Zhen;Nie, Bihua;Zhang, Jiang

作者机构：

关键词： Cas13a; Guide RNA; Multiplex RNA targeting; Potato virus Y; Resistance

期刊名称：PLANTA （ 影响因子：4.3； 五年影响因子：4.8 ）

ISSN： 0032-0935

年卷期： 2023 年 258 卷 4 期

页码：

收录情况： SCI

摘要： Main conclusionThe Cas13a-based multiplex RNA targeting system can be engineered to confer resistance to RNA viruses, whereas the number and expression levels of gRNAs have no significant effect on viral interference.AbstractThe CRISPR-Cas systems provide adaptive immunity to bacterial and archaeal species against invading phages and foreign plasmids. The class 2 type VI CRISPR/Cas effector Cas13a has been harnessed to confer the protection against RNA viruses in diverse eukaryotic species. However, whether the number and expression levels of guide RNAs (gRNAs) have effects on the efficiency of RNA virus inhibition is unknown. Here, we repurpose CRISPR/Cas13a in combination with an endogenous tRNA-processing system (polycistronic tRNA-gRNA) to target four genes of potato virus Y (PVY) with varying expression levels. We expressed Cas13a and four different gRNAs in potato lines, and the transgenic plants expressing multiple gRNAs displayed similar suppression of PVY accumulation and reduced disease symptoms as those expressing a single gRNA. Moreover, PTG/Cas13a-transformed plants with different expression levels of multiple gRNAs displayed similar resistance to PVY strains. Collectively, this study suggests that the Cas13a-based multiplex RNA targeting system can be utilized to engineer resistance to RNA viruses in plants, whereas the number and expression levels of gRNAs have no significant effect on CRISPR/Cas13a-mediated viral interference in plants.

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