Antiviral efficacy of short-hairpin RNAs and artificial microRNAs targeting foot-and-mouth disease virus

• Published in: PeerJ (San Francisco, CA) Vol. 9; p. e11227
• Main Authors: Currá, Anabella, Cacciabue, Marco, Gravisaco, María José, Asurmendi, Sebastián, Taboga, Oscar, Gismondi, María I
• Format: Journal Article
• Language: English
• Published: United States PeerJ. Ltd 09.06.2021; PeerJ, Inc; PeerJ Inc
• Subjects: Animal diseases; Antiviral activity; Antiviral agents; Antivirals; Biotechnology; Cell Biology; Cell lines; DNA-directed RNA polymerase; Epidemiology; Ethylenediaminetetraacetic acid; FMDV; Foot & mouth disease; Foot-and-mouth disease; Gene silencing; Genes; Genetic engineering; Genetic transcription; MicroRNA; MicroRNAs; miRNA; Molecular Biology; Nucleotide sequence; Plasmids; Post-transcription; Replication; RNA interference; RNA viruses; RNA-directed RNA polymerase; RNA-mediated interference; Target accessibility; Vaccination; Veterinary Medicine; Virology; Viruses; United States > US; Argentina; FMDV; Target accessibility; RNA interference; Antivirals
• ISSN: 2167-8359; 2167-8359
• DOI: 10.7717/peerj.11227

RNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directs post-transcriptional gene silencing through small RNA molecules. RNAi has been proposed as an alternative approach for rapid and specific control of viruses including foot-and-mouth disease virus (FMDV), the causative agent of a devastating animal disease with high economic impact. The aim of this work was to assess the antiviral activity of different small RNA shuttles targeting the FMDV RNA-dependent RNA polymerase coding sequence (3D). Three target sequences were predicted within 3D considering RNA accessibility as a major criterion. The silencing efficacy of short-hairpin RNAs (shRNAs) and artificial microRNAs (amiRNAs) targeting the selected sequences was confirmed in fluorescent reporter assays. Furthermore, BHK-21 cells transiently expressing shRNAs or amiRNAs proved 70 to >95% inhibition of FMDV growth. Interestingly, dual expression of amiRNAs did not improve FMDV silencing. Lastly, stable cell lines constitutively expressing amiRNAs were established and characterized in terms of antiviral activity against FMDV. As expected, viral replication in these cell lines was delayed. These results show that the target RNA-accessibility-guided approach for RNAi design rendered efficient amiRNAs that constrain FMDV replication. The application of amiRNAs to complement FMDV vaccination in specific epidemiological scenarios shall be explored further.