Inhibition of foot-and-mouth disease virus replication by small interfering RNA

Abstract

Foot-and-mouth disease, caused by foot-and-mouth disease virus (FMDV), is one of the most dangerous diseases of cloven-hoofed animals and is a constant threat to the dairy and beef industries in the Middle East and other regions of the world, despite intensive vaccination programmes. In this work, the ability of specific small interfering (si)RNAs to inhibit virus replication in BHK-21 cells was examined. By using bioinformatic computer programs, all FMDV sequences in public-domain databases were analysed. The analysis revealed three regions of at least 22 bp with 100 % identity in all FMDV entries. From these sequences, three specific siRNA molecules were prepared and used to test the ability of siRNAs to inhibit virus replication. By using real-time quantitative PCR to measure the amount of viral RNA in infected cells, it was shown that virus replication was inhibited in cells that were transfected with siRNAs. When viral titres were examined, 100 % inhibition of growth could be demonstrated in cells transfected with a mixture of all three anti-FMDV siRNAs, compared with control cells transfected with anti-LacZ siRNA.

Inhibition of Foot-and-Mouth Disease Virus Infections in Cell Cultures with Antisense Morpholino Oligomers

“…Their study showed that microinjection of antisense DNA or RNA targeting the AUG2 region into the cytoplasms of BHK-21 cells caused transient inhibition of FMDV infections. The magnitude of viral inhibition produced by FMDV-specific PPMO compares quite favorably with results from studies with other nucleic-acid-based agents evaluated against FMDV, such as small interfering RNA (8,10,19,21,25), antisense DNA (18), and antisense RNA (3,17).…”

“…Sense and antisense RNAs (3,17,18,34), antisense DNA (18), and small interfering RNA (8,10,19) have generated various levels of antiviral activity. All of these strategies apparently function by interfering with expression of the viral RNA genome.…”

A Morpholino Oligomer Targeting Highly Conserved Internal Ribosome Entry Site Sequence Is Able To Inhibit Multiple Species of Picornavirus

“…A number of compounds designed to inhibit picornavirus infections through an antisense-mediated mechanism have generated positive preclinical results but are still early in the development process. Antisense phophorothioate DNA has been used against coxsackievirus type B3 (CVB3) in vitro (57) and in vivo (62), antisense RNA has been used against foot-and-mouth disease virus (45), and small interfering RNA has been used against several picornaviruses (3,20,21,26,42,47), all with some success. Phosphorodiamidate morpholino oligomers (PMO) are composed of individual subunits consisting of a DNA base connected to a morpholine ring and phophorodiamidate-linked backbone (50).…”

Inhibition of viral replication by small interfering RNA targeting of the foot-and-mouth disease virus receptor integrin β6

“…RNAi is an effective method for inhibiting viral replication, and previous studies have investigated the effects of anti-viral RNAi methods on FMDV (29,30). The initial stage of host cell infection by FMDV involves interaction with a cognate cell-surface receptor, which subsequently enables virus particles to enter cells in a receptor-mediated manner.…”