Establishing RNA virus resistance in plants by harnessing CRISPR immune system

Recently, CRISPR-Cas (clustered, regularly interspaced short palindromic repeats–CRISPR-associated proteins; 成簇恒间隔短回文重复序列-CRISPR-相关蛋白) system has been used to produce plants resistant to DNA virus infections. However, there is no RNA virus control method in plants that uses CRISPR-Cas system to target the viral genome directly. Here, we reprogrammed the CRISPR-Cas9 system from Francisella novicida to confer molecular immunity against RNA viruses in Nicotiana benthamiana (本氏烟) and Arabidopsis plants. Plants expressing FnCas9 and sgRNA specific for the cucumber mosaic virus (CMV; 黄瓜花叶病毒) or tobacco mosaic virus (TMV; 烟草花叶病毒) exhibited significantly attenuated (减弱) virus infection symptoms (症状) and reduced viral RNA accumulation. Furthermore, in the transgenic virus-targeting plants, the resistance was inheritable and the progenies showed significantly less virus accumulation. These data reveal that the CRISPR/Cas9 system can be used to produce plant that stable resistant to RNA viruses, thereby broadening (拓宽) the use of such technology for virus control in agricultural field.

近来成簇恒间隔短回文重复序列-CRISPR-相关蛋白，即CRISPR-Cas系统被用于赋予植物对于DNA病毒侵染的抗性。然而，并未有针对RNA病毒的CRISPR-Cas系统被开发出来。本文报道了一种重编程的CRISPR-Cas9，可以赋予烟草和拟南芥对于RNA病毒Francisella novicida的分子免疫。在植株中表达FnCas9和特异的黄瓜花叶病毒或是烟草花叶病毒的sgRNA，转基因植株的病毒侵染症状显著减轻，RNA病毒的积累也有所减少。此外，在转基因植株中，抗性可以遗传，子代表现出更少的病毒积累特征。本文的结果显示CRISPR/Cas9系统可以被用来辅助植物提升RNA病毒抗性，因此拓宽了该技术在农业领域的病毒防治应用。

（p.s. 2014.9.28加州大学伯克利分校的Jennifer Doudna实验室在Nature上报道了一种可以重编程CRISPR/Cas9蛋白复合物在序列特异性的靶位点识别并切割RNA的方法；2015.4.27埃默里大学的研究人员在PNAS上报道了利用CRISPR-Cas赋予了人类细胞针对Francisella novicida的抗性的研究）

通讯：周国辉 (http://nxy.scau.edu.cn/2016/1209/c581a18613/page.htm)

个人简介：1984年，安徽农学院植物保护专业，学士；1988年，安徽农业大学植物病理学，硕士；1997-2000年，华南农业大学，博士。

doi: 10.1111/pbi.12881