Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers
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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.
Forrest , S , Lear , Z , Herod , M R , Ryan , M , Rowlands , D J & Stonehouse , N J 2014 , ' Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers ' Journal of General Virology , vol 95 , pp. 2649-2657 . DOI: 10.1099/vir.0.067751-0
Journal of General Virology
Copyright 2014 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Sophie Forrest was funded by a BBSRC studentship and by the Wellcome Trust (094898). Zoe Lear is funded by a Wellcome Trust 4-year PhD programme (The Molecular Basis of Biological Mechanisms).