Effective inhibition of foot-and-mouth disease virus (FMDV) replication in vitro by vector-delivered microRNAs targeting the 3D gene

Abstract

BackgroundFoot-and-mouth disease virus (FMDV) causes an economically important and highly contagious disease of cloven-hoofed animals. RNAi triggered by small RNA molecules, including siRNAs and miRNAs, offers a new approach for controlling viral infections. There is no report available for FMDV inhibition by vector-delivered miRNA, although miRNA is believed to have more potential than siRNA. In this study, the inhibitory effects of vector-delivered miRNAs targeting the 3D gene on FMDV replication were examined.ResultsFour pairs of oligonucleotides encoding 3D-specific miRNA of FMDV were designed and selected for construction of miRNA expression plasmids. In the reporter assays, two of four miRNA expression plasmids were able to significantly silence the expression of 3D-GFP fusion proteins from the reporter plasmid, p3D-GFP, which was cotransfected with each miRNA expression plasmid. After detecting the silencing effects of the reporter genes, the inhibitory effects of FMDV replication were determined in the miRNA expression plasmid-transfected and FMDV-infected cells. Virus titration and real-time RT-PCR assays showed that the p3D715-miR and p3D983-miR plasmids were able to potently inhibit the replication of FMDV when BHK-21 cells were infected with FMDV.ConclusionOur results indicated that vector-delivered miRNAs targeting the 3D gene efficiently inhibits FMDV replication in vitro. This finding provides evidence that miRNAs could be used as a potential tool against FMDV infection.

Multiple microRNAs targeted to internal ribosome entry site against foot-and-mouth disease virus infection in vitro and in vivo

“…Each miRNA expression plasmid (including pmiR-NC, the negative control; Table 1) was, with each reporter plasmid (including p3D-GFP, a control for nonspecific effects [41]), used to co-transfected BHK-21 cells at a molar ratio of 1:1 (w/w). The cells were observed continuously under the fluorescence microscope and analyzed by flow cytometry 48 h post-transfection.…”

MicroRNA-23b Inhibits Enterovirus 71 Replication through Downregulation of EV71 VPl Protein

“…Previous studies have also provided supportive proof of this conclusion. For instance, the results from Du et al [19] showed that vector-delivered miRNAs targeting the 3D genes efficiently inhibited the FMDV replication in vitro. Ahluwalia et al [20] found that human cellular miRNA hsa-miR-29a interfered with the expression of viral Nef protein and HIV-1 replication.…”

In Vivo, Cardiac-Specific Knockdown of a Target Protein, Malic Enzyme- 1, in Rat via Adenoviral Delivery of DNA for Non-Native miRNA

“…As with shRNA, delivery of non-native miRNA constructs have been used to suppress target protein expression in cell cultures [4-6]. In whole animal models [30,31], pioneering studies in the cancer field have described impressive therapeutic effects with these non-native miRNAs, yet the approach had not been tested in heart, in vivo .…”

“…With the recent developments in RNA interference technologies (RNAi), it is highly anticipated that these viral-based delivery approaches can be extended for the suppression of target protein expression in the heart [3]. The approach has been overwhelmingly successful in isolated cell culture models [4-6], including several studies for the suppression of malic enzyme as targeted in this study [7-9]. However, there are only a few reports for successful suppression of a target protein in heart, in vivo , based on RNAi schemes [10-14].…”