In this study, we show that 15 nm citrate-capped AuNPs exert a remarkable toxicity in living systems. The assessment was performed by using well-characterized AuNPs, the combination of in vitro and in vivo models (namely two different cell lines and Drosophila melanogaster), exposure to low dosages of nanoparticles (in the sub-nanomolar concentration range), along with the application of several biological assays to monitor different aspects of the toxic effects, such as viability, genotoxicity, and molecular biomarkers.