Gene silencing is an important tool for increasing the yield of crops and plays an important role in revolutionizing agriculture. In agriculture, gene silencing is done by post-transcriptional gene silencing (PTGS) and transcriptional gene silencing (TGS) technology. RNA interference (RNAi) is a well-known technique of post-transcriptional gene silencing and is widely utilized in crop improvement through gene regulation. In the present review, we discussed various aspects of RNA interference gene silencing viz RNA-dependent RNA polymerase (RdRp), Argonaute proteins (AGOs) and Dicer-like proteins (DCLs) and their applications in the optimization of agricultural yield.