Abstract:
The intrinsic nature of CRISPR-Cas in conferring immunity to bacteria and archaea has been repurposed to combat pathogenic agents in mammalian and plant cells. In this regard, CRISPR-Cas13 systems have proved their remarkable potential for single-strand RNA viruses targeting. Here, different types of Cas13 orthologs were applied to knockdown foot-and-mouth disease virus (FMDV), a highly contagious disease of a wide variety of species with genetically diverse strains and is widely geographically distributed. Using programmable CRISPR RNAs capable of targeting conserved regions of the viral genome, all Cas13s from CRISPR system type VI (subtype A/B/D) could comprehensively target and repress different serotypes of FMDV virus. This approach has the potential to destroy all strains of a virus as targets the ultra-conserved regions of genome. We experimentally compared the silencing efficiency of CRISPR and RNAi by designing the most effective short hairpin RNAs according to our developed scoring system and observed comparable results. This study showed successful usage of various Cas13 enzymes for suppression of FMDV, which provides a flexible strategy to battle with other animal infectious RNA viruses, an underdeveloped field in the biotechnology scope.
摘要:
CRISPR-Cas在赋予细菌和古细菌免疫力方面的固有性质已被重新用于对抗哺乳动物和植物细胞中的病原体。在这方面,CRISPR-Cas13系统已经证明了它们在单链RNA病毒靶向方面的巨大潜力。这里,将不同类型的Cas13直向同源物应用于击倒口蹄疫病毒(FMDV),一种种类繁多的高度传染性疾病,具有遗传多样性的菌株,并且在地理上分布广泛。使用能够靶向病毒基因组保守区的可编程CRISPRRNA,来自CRISPR系统VI型(A/B/D亚型)的所有Cas13s都可以全面靶向和抑制不同血清型的FMDV病毒。这种方法具有破坏作为基因组超保守区域靶标的所有病毒株的潜力。我们通过根据我们开发的评分系统设计最有效的短发夹RNA,实验比较了CRISPR和RNAi的沉默效率,并观察到了可比的结果。这项研究表明,成功使用各种Cas13酶抑制FMDV,这提供了一个灵活的策略来对抗其他动物传染性RNA病毒,生物技术领域的不发达领域。