Fluorometric RdRp assay with self-priming RNA
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.
KeywordsViral RdRp RNA-dependent RNA polymerase Fluorometric RdRp assays dsRNA and ssRNA
This study was funded by North American University, Houston, Texas, and Yeditepe University, Istanbul.
Conflict of interest
All authors declare that they have no conflicts of interest concerning this work.