Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers

S. Forrest, Z. Lear, M.R. Herod, M. Ryan, D.J. Rowlands, N.J. Stonehouse

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Abstract

We have previously documented the inhibitory activity of RNA aptamers to the RNA-dependent RNA polymerase of foot-and-mouth disease virus (3D). Here we report their modification and use with a subgenomic replicon incorporating GFP (pGFP-PAC replicon), allowing replication to be monitored and quantified in real-time. GFP expression in transfected BHK-21 cells reached a maximum at approximately 8 h post-transfection, at which time change in morphology of the cells was consistent with a virus-induced cytopathic effect. However, transfection of replicon-bearing cells with a 3D aptamer RNA resulted in inhibition of GFP expression and maintenance of normal cell morphology, whereas a control aptamer RNA had little effect. The inhibition was correlated with a reduction in 3D (detected by immunoblotting) and shown to be dose dependent. The 3D aptamers appeared to be more effective than 29-C-methylcytidine (29CMC). Aptamers to components of the replication complex are therefore useful molecular tools for studying viral replication and also have potential as diagnostic molecules in the future.
Original languageEnglish
Pages (from-to)2649-2657
Number of pages9
JournalJournal of General Virology
Volume95
Early online date5 Aug 2014
DOIs
Publication statusPublished - 1 Dec 2014

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