RNA interference (RNAi) is an endogenous gene regulation mechanism that exists in nearly all eukaryotic organisms. Various versions of RNAi have been used in biotechnology to manipulate gene expression in many plant species. Exogenous application of double-stranded RNA (dsRNA) to induce RNAi represents sustainable alternatives to traditional transgenic, breeding or chemical-based crop protection and trait modification strategies. While all eukaryotic organisms, a priori contain a functional RNAi pathway not all are effective targets of exogenously applied dsRNA. The two major factors that dictate whether an organism is an effective target for exogenous RNAi are the ability to take up trigger dsRNA and the ability of the organism to process this into functional siRNAs. We have identified a compound that allows uptake of dsRNA into plant cells to initiate highly effective gene silencing and virus protection. Not only was enhanced cellular uptake seen for RNA but also for other macromolecules including protein and nanoparticles. Beyond this we have utilised a novel small RNA purification protocol that has allowed us to diagnose functional siRNAs produced after cellular uptake of initiating RNA molecules. Our mechanistic dissection of the uptake and processing of exogenous RNAi provides crucial insights into this novel technology and stands to add significant benefit to this emerging field of plant protection and trait modification.