title: Inhibition of the foot-and-mouth disease virus subgenomic replicon by RNA aptamers.
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.
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Sophie
authors.initials: S
authors.lastname: Forrest
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Zoe
authors.initials: Z
authors.lastname: Lear
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Morgan R
authors.initials: MR
authors.lastname: Herod
authors.affiliation: Biomedical Sciences Research Complex (BSRC), School of Biology, University of St Andrews, North Haugh, St Andrews, Fife KY16 9ST, UK.
authors.forename: Martin
authors.initials: M
authors.lastname: Ryan
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Rowlands
authors.affiliation: School of Molecular and Cellular Biology, Faculty of Biological Sciences and Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds LS2 9JT, UK.
authors.forename: Nicola J
authors.initials: NJ
authors.lastname: Stonehouse
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide; D002460:Cell Line; D006224:Cricetinae; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000502:physiology*; D015967:Gene Expression Regulation, Viral / Q000502:physiology; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics; D016296:Mutagenesis; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D012093:Replicon
chemical_list: D052157:Aptamers, Nucleotide; D008164:Luminescent Proteins; D012367:RNA, Viral; D049452:Green Fluorescent Proteins
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