Co-precipitation, a minimal technique, was used successfully to create a novel (Cr, Cu) dual-doped ZnO nanoparticles (NPs). X-ray diffraction (XRD), energy dispersion X-ray (EDX) analysis, scanning electron microscopes (SEM), and UV–visible absorption spectra had been used to characterize the produced nanoparticle. According to the XRD pattern, the produced NPs have a hexagonal wurtzite structure with a high phase purity. The substitution of Cr increased optical absorption and induced new absorption peaks in the visible regions that correspond to the transitions from 4A2(4F) to CB and 4A2(4F) to 4T1(4F). Absorption of photons in the visible region led to an increase in the photo-generated electron–hole pairs and hence RO-species which are responsible for photocatalytic activity. Therefore, the (Cu, Cr) dual doped ZnO NPs had unique photocatalytic activity under sunlight irradiation. MTT assay was employed to analyse the in-vitro cytotoxicity of Cr-doped Zn0.98Cu0.02O NPs on human normal PBMC. In which, the toxicity is enhanced by a proliferation in Cr concentration and has very small amounts at low concentrations and significantly at high concentrations. The antibacterial characteristics of the synthesized NPs were examined against the bacterial strains P. putida and B. subtilis.