Extracellular vesicles produced by the protozoan parasite Trichomonas vaginalis contain a preferential cargo of tRNA-derived small RNAs

[Display omitted] •Trichomonas vaginalis extracellular vesicles (EVs) encapsulated RNAs of small size.•This cargo was predominantly composed of tRNA-derived small RNAs (tsRNAs), an emerging class of small regulatory RNAs.•Specificity of the tsRNA cargo, with preference for 5′ tRNA halves, indicates a selective process for EV packaging.•The EV RNA cargo of T. vaginalis was actively internalised by human cells. Trichomonas vaginalis is a protozoan parasite that causes trichomoniasis, the most prevalent non-viral sexually transmitted infection worldwide. Trichomonas vaginalis releases extracellular vesicles that play a role in parasite:parasite and parasite:host interactions. The aim of this study was to characterise the RNA cargo of these vesicles. Trichomonas vaginalis extracellular vesicles were found to encapsulate a cargo of RNAs of small size. RNA-seq analysis showed that tRNA-derived small RNAs, mostly 5′ tRNA halves, are the main type of small RNA in these vesicles. The tRNA-derived small RNAs in T. vaginalis extracellular vesicles were shown to be derived from the specific processing of tRNAs within cells. The specificity of this RNA cargo in T. vaginalis extracellular vesicles suggests a preference for packaging. The RNA cargo of T. vaginalis was shown to be rapidly internalised by human cells via lipid raft-dependent endocytosis. The potential role of these tsRNAs – an emerging class of small RNAs with regulatory functions – on altering host cellular responses requires further examination, suggesting a new mode of parasite:host communication.