Efficient inhibition of foot-and-mouth disease virus replication in vitro by artificial microRNA targeting 3D polymerase

Abstract

Foot-and-mouth disease (FMD) is a devastating acute viral disease of livestock with cloven hooves. Among various therapeutic control measures, RNA interference (RNAi) is one of the methods being explored to inhibit FMD virus replication and spread. The RNAi is achieved by short hairpin RNAs or artificial microRNAs (amiRNAs). Utility of amiRNAs as antiviral, targeting conserved regions of the viral genome is gaining importance. However, delivery of miRNA in vivo is still a challenge. In this study, the efficacy of amiRNAs in preventing FMD virus replication in a permissive cell culture system was investigated, by generating stable cell lines expressing amiRNAs targeting three functional regions of the FMD virus (FMDV) genome (IRES, 3B3 and 3D). The results showed that amiRNA targeting 3D polymerase is relatively more efficient. However, expression of multiple microRNAs targeting the three regions did not exhibit additive effect. The data suggest that 3D specific miRNA is a potential valid strategy in developing novel antiviral measures against FMDV infection.

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

“…As shown in the reporter assays, the predicted targets were silenced efficiently independently of the RNAi mediator used, supporting the feasibility of the algorithm applied for target selection. Our results are in accordance with previous data indicating that the 3D region of the FMDV genome is a suitable target for RNAi ( Pengyan et al, 2008 ; Du et al, 2011 ; Gu et al, 2014 ; Basagoudanavar et al, 2019 ) and they represent a step forward in the identification of new target sequences within FMDV, specifically in serotype A, that had been less explored in previous works.…”

“…To overcome this drawback, artificial miRNAs (i.e., miRNAs rationally designed to target defined sequences) have been favored as RNAi shuttles since they exhibit increased safety with comparable efficacy ( Boudreau, Martins & Davidson, 2009 ). The efficacy of artificial miRNAs (amiRNAs) against FMDV has been investigated to a lesser extent ( Du et al, 2011 ; Gismondi et al, 2014 ; Basagoudanavar et al, 2019 ). In this work, we further assess the efficacy of amiRNAs against FMDV and shRNAs targeting the same viral regions in susceptible cells.…”

E2 ubiquitin-conjugating enzyme UBE2L6 promotes Senecavirus A proliferation by stabilizing the viral RNA polymerase

“…The 3D protein of SVA is an RNA-dependent RNA polymerase, as in other picornaviruses it is essential for viral replication [ 37 ]. The microRNAs targeting the 3D polymerase of FMDV effectively inhibit virus replication in vitro [ 38 ]. Monoclonal antibodies targeting the 3D protein of enterovirus 71 (EV71) affect its RdRp activity and inhibit virus replication [ 39 ].…”

MiR-4334-5p Facilitates Foot and Mouth Disease Virus Propagation by Suppressing Interferon Pathways via Direct Targeting ID1

“…For instance, miR-101 represses feline herpesvirus 1 reproduction by targeting cellular suppressor of cytokine signaling 5 to promote IFN-I signaling [ 20 ]; miR-7 suppresses rotavirus replication via targeting viral NSP5 directly [ 21 ]; miR-122 reduces HBV gene expression and replication via binding with hepatitis B virus pre-genomic RNA sequence [ 22 ]; miR-21-3p promotes influenza A virus H5N1 replication by regulating FGF2 to repress IFN-I signaling [ 23 ]; miR-545 promotes enterovirus 71 propagation through directly targeting phosphatase and tensin homolog and tumor necrosis factor receptor-associated factor 6 [ 24 ]. In the case of FMDV, there are only a few studies, artificial miRNA targeting 3D polymerase is efficient to inhibit FMDV replication [ 25 ], miR-1307 and miR-203a repress FMDV infection [ 2 , 15 ]. However, the underlying mechanisms of porcine miR-4334-5p affecting viral replication are still not understood.…”

“…The disease mainly occurred in cloven-hoofed animals, such as cattle, sheep, goats, and swine, clinical symptoms in infected animals usually characterized as the vesicles formation in the mouth, nose, tongue and skin between the claws of the feet [ 26 ]. There were multiple artificial designed miRNAs targeting viral 3D pol or internal ribosome entry site, which had been proved to impair FMDV propagation significantly [ 25 ]. In addition, miRNA-based treatment of Hepatitis C infection has successfully completed phase II clinical trial [ 27 ].…”