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Mass sequencing of cDNA libraries from salivary glands of triatomines has resulted in the identification of many novel genes of unknown function. The aim of the present work was to develop a functional RNA interference (RNAi) technique for Rhodnius prolixus, which could be widely used for functional genomics studies in triatomine bugs. To this end, we investigated whether double-stranded RNA (dsRNA) can inhibit gene expression of R. prolixus salivary nitrophorin 2 (NP2) and what impact this might have on anticoagulant and apyrase activity in the saliva. dsRNA was introduced by two injections or by ingestion. RT-PCR of the salivary glands showed that injections of 15μg of NP2 dsRNA in fourth-instar nymphs reduced gene expression by 75±14% and that feeding 1μg/μL of NP2 dsRNA into second-instar nymphs (approx. 13μg in total) reduced gene expression by 42±10%. Phenotype analysis showed that …