Unfolding of in planta activity of anti‐rep ribozyme in presence of a RNA silencing suppressor

Mishra, Sumona Karjee; Chilakamarthi, Ushasri; Deb, J.K.; Mukherjee, Sunil Kumar

Antisense RNA ribozymes have intrinsic endonucleolytic activity to effect cleavage of the target RNA. However, this activity in vivo is often controlled by the dominance of antisense or other double‐stranded RNA mechanism. In this work, we demonstrate the in planta activity of a hammerhead ribozyme designed to target rep‐mRNA of a phytopathogen Mungbean Yellow Mosaic India virus (MYMIV) as an antiviral agent. We also found RNA‐silencing is induced on introduction of catalytically active as well as inactive ribozymes. Using RNA‐silencing suppressors (RSS), we demonstrate that the endonucleolytic activity of ribozymes is a true phenomenon, even while a mutated version may demonstrate a similar down‐regulation of the target RNA. This helps to ease the confusion over the action mechanism of ribozymes in vivo.