Abiotic and biotic stresses severely affect plant growth and yield. However, plants can adapt to biotic stresses by reprogramming gene expression at the transcriptional and post-transcriptional levels. Recently, the exogenous application of RNA interference (RNAi) technology is making good progress and offering a possible alternative to genetic transformation for combating biotic stresses in plants. In this process, the RNAi regulates the transcript level by transcriptional gene silencing (suppression transcription) or post-transcriptional gene silencing (activating sequence-specific RNA degradation). The use of RNAi tools and their respective targets against various biotic stresses has been reported in many agronomic crops. Spray-induced gene silencing is a recent advancement that uses dsRNA as a trigger to silence target genes in pathogens, however, the carrier is needed to stabilise the RNA for a longer period and to reach the target pathogen. In this review, we focus on the synthesis of RNAi constructs and their potential applications against biotic stressors in different crop species.