Forrest, S, Lear, Z, Herod, M et al. (3 more authors) (2014) Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers. Journal of General Virology, 95 (Pt 12). 2649 - 2657. ISSN 0022-1317
Abstract
We have previously documented the inhibitory activity of RNA aptamers to the RNA-dependent RNA polymerase of foot-and-mouth disease virus (3D(pol)). 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(pol) 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(pol) (detected by immunoblotting) and shown to be dose dependent. The 3D(pol) aptamers appeared to be more effective than 2'-C-methylcytidine (2'CMC). 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.
Metadata
Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2014, The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
Keywords: | Animals; Aptamers, Nucleotide; Cell Line; Cricetinae; Foot-and-Mouth Disease Virus; Gene Expression Regulation, Viral; Genetic Engineering; Green Fluorescent Proteins; Luminescent Proteins; Mutagenesis; RNA, Viral; Replicon |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 25 Mar 2015 10:55 |
Last Modified: | 17 Aug 2015 13:36 |
Published Version: | http://dx.doi.org/10.1099/vir.0.067751-0 |
Status: | Published |
Publisher: | Society for General Microbiology |
Identification Number: | https://doi.org/10.1099/vir.0.067751-0 |
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