Virus Genes

, Volume 50, Issue 3, pp 498–504 | Cite as

Fluorometric RdRp assay with self-priming RNA

Article

Abstract

There is an outmost need for the identification of specific antiviral compounds. Current antivirals lack specificity, making them susceptible to off-target effects, and highlighting importance of development of assays to discover antivirals targeting viral specific proteins. Previous studies for identification of inhibitors of RNA-dependent RNA polymerase (RdRp) mostly relied on radioactive methods. This study describes a fluorometric approach to assess in vitro activity of viral RdRp for drug screening. Using readily available DNA- and RNA-specific fluorophores, we determined an optimum fluorometric approach that could be used in antiviral discovery specifically for RNA viruses by targeting RdRp. Here, we show that double-stranded RNA could be successfully distinguished from single-stranded RNA. In addition, we provide a strategy based on self-priming RNA to assess RdRp activity.

Graphical abstract

Keywords

Viral RdRp RNA-dependent RNA polymerase Fluorometric RdRp assays dsRNA and ssRNA 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Fatih Kocabas
    • 1
    • 2
  • Raife D. Turan
    • 1
  • Galip S. Aslan
    • 1
  1. 1.Department of Genetics and Bioengineering, Faculty of EngineeringYeditepe UniversityIstanbulTurkey
  2. 2.Department of EducationNorth American UniversityHoustonUSA

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