Efficient RNA Virus Targeting via CRISPR/CasRx in Fish
Qing Wang, Yun Liu, Chong Han, Min Yang, Fengqi Huang, Xuzhuo Duan, Shaowen Wang, Yepin Yu, Jiaxin Liu, Huirong Yang, Danqi Lu, Huihong Zhao, Yong Zhang, Qiwei Qin
Abstract
RNA viruses are important viral pathogens infecting vertebrates and mammals. RNA virus populations are highly dynamic due to short generation times, large population sizes, and high mutation frequencies. Therefore, it is difficult to find widely effective ways to inhibit RNA viruses, and we urgently need to develop effective antiviral methods. CasRx is a small type VI-D effector (Cas13d) with RNA knockdown efficiency that can have an interference effect on RNA viruses. Nervous necrosis virus (NNV), a nonenveloped positive-strand RNA virus, is one of the most serious viral pathogens, infecting more than 40 cultured fish species and resulting in huge economic losses worldwide. Here, we establish a novel effective CasRx system for RNA virus interference using NNV and grouper (Epinephelus coioides) as a model. Our data showed that CasRx was most robust for RNA virus interference applications in fish, and we demonstrate its suitability for studying key questions related to virus biology.