Asadbeigi Adnan, Bakhtiarizadeh Mohammad Reza, Saffari Mojtaba, Modarressi Mohammad Hossein, Sadri Naser, Kafi Zahra Ziafati, Fazilaty Hassan, Ghalyanchilangeroudi Arash, Esmaeili Hossein
Cancer Institute, Department of Medical Genetics, Faculty of Medicine, Tehran University of Medical Sciences (TUMS), Tehran 1417613151, Iran.
Department of Animal and Poultry Science, College of Aburaihan, University of Tehran, Tehran 3391653755, Iran.
Mol Ther Nucleic Acids. 2024 May 31;35(3):102235. doi: 10.1016/j.omtn.2024.102235. eCollection 2024 Sep 10.
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.
CRISPR-Cas赋予细菌和古细菌免疫的内在特性已被重新利用，用于对抗哺乳动物和植物细胞中的病原体。在这方面，CRISPR-Cas13系统已证明其在靶向单链RNA病毒方面具有显著潜力。在此，不同类型的Cas13直系同源物被用于敲低口蹄疫病毒（FMDV），这是一种具有高度传染性的疾病，可感染多种物种，病毒株具有遗传多样性且在地理上广泛分布。使用能够靶向病毒基因组保守区域的可编程CRISPR RNA，来自CRISPR系统VI型（A/B/D亚型）的所有Cas13都可以全面靶向并抑制不同血清型的FMDV病毒。这种方法有可能摧毁作为靶向基因组超保守区域的病毒的所有毒株。我们根据开发的评分系统设计了最有效的短发夹RNA，通过实验比较了CRISPR和RNAi的沉默效率，并观察到了类似的结果。这项研究表明各种Cas13酶成功用于抑制FMDV，这为对抗其他动物传染性RNA病毒提供了一种灵活策略，而这在生物技术领域是一个尚未充分发展的领域。
