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

ANABELLA P. CURRÁ; CACCIABUE. MARCO; GRAVISACO, MARIA JOSE; ASURMENDI SEBASTIAN; OSCAR ALBERTO TABOGA; GISMONDI MI

PeerJ

PeerJ

Año: 2021

RNA interference (RNAi) is a well-conserved mechanism in eukaryotic cells that directsposttranscriptional gene silencing through small RNA molecules. RNAi has been proposedas an alternative approach for rapid and specific control of viruses including foot-andmouthdisease virus (FMDV), the causative agent of a devastating animal disease with higheconomic impact. The aim of this work was to assess the antiviral activity of different smallRNA shuttles targeting the FMDV RNA-dependent RNA polymerase coding sequence (3D).Three target sequences were predicted within 3D considering RNA accessibility as a majorcriterion. 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 improveFMDV silencing. Lastly, stable cell lines constitutively expressing amiRNAs wereestablished 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 RNAaccessibility-guided approach for RNAi design rendered efficient amiRNAs that constrainFMDV replication. The application of amiRNAs to complement FMDV vaccination in specificepidemiological scenarios shall be explored further.