Bilsen, A.; Ottati, S.; Parise, C.; Abba, S.; Vanden Broeck, J.; Galetto, L.; Santos, D.

Growing evidence shows that eukaryotic genomes contain DNA sequences of non-retroviral RNA virus origin, yet the mechanisms underlying the generation of this virus-derived complementary DNA (vDNA) remain poorly understood, particularly in insects. Here, we demonstrate that infection with diverse non-retroviral RNA viruses induces the production of reverse-transcribed vDNA across multiple lepidopteran and hemipteran species. We further show that a Drosophila melanogaster-derived cell line actively secretes reverse-transcribed vDNA associated with extracellular vesicles (EVs), widely conserved nanoscale mediators of intercellular communication. Together, these findings reveal that infection-induced vDNA formation is a widespread phenomenon in insects and suggest that EVs may facilitate the systemic dissemination of vDNA.