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.
Department of Genetics and Bioengineering, Faculty of Engineering, Yeditepe University, 34755, Istanbul, Turkey, fatih.kocabas@yeditepe.edu.tr.
Fatih
F
Kocabas
Raife D
RD
Turan
Galip S
GS
Aslan
D000998:Antiviral Agents / Q000302:isolation & purification* / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005470:Fluorometry / Q000379:methods*; D012324:RNA-Dependent RNA Polymerase / Q000032:analysis* / Q000037:antagonists & inhibitors*
D000998:Antiviral Agents; D012324:RNA-Dependent RNA Polymerase