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

CURRÁ, ANABELLA; CACCIABUE, MARCO; GRAVISACO, MARÍA JOSÉ; ASURMENDI, SEBASTIÁN; TABOGA, OSCAR; GISMONDI, MARÍA I.

PeerJ

PeerJ Inc.

Año: 2021 vol. 9

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