Metal oxide nanostructures have shown significant promise for biosensors, gas sensors, photocatalyst and other biomedical applications. Among these, zinc oxide (ZnO) nanostructures, exhibiting interesting properties such as high catalytic activity, biocompatibility, high isoelectric point, large surface to volume ratio, make them a good candidate for biosensing applications. Here we report the synthesis of ZnO nanorods (ZnONR) on ITO films in aqueous phase and its application in Urea biosensor fabrication. ZnONR have been synthesized by a two-step method, first seed growth of ZnO by sputtering on ITO films followed by decomposition of zinc nitrate hexahydrate / hexamethylenetetramine (HMT) in aqueous phase. Exploiting the high isoelectric point of ZnO, a Urease/ZnONR/ITO bioelectrode has been fabricated by physical binding of Urease (Urs) onto ZnONRs. X-ray diffraction (XRD), field emission-scanning electron microscopy (FE-SEM), and cyclic voltammetry (CV) have been used to characterize ZnONR and the Urs/ZnONR/ITO bioelectrode. The FE-SEM and XRD measurements confirm the formation of ZnONR. The electrochemical data from the Urs/ZnONR/ITO biolectrode reveal linearity between 1-11 mM with sensitivity of 0.9 μA/mM and a relatively low Michaelis-Menten constant (Km) of 5.01 mM for urea sensing. The results indicate the potential of ZnONR films for fabrication of commercial biosensors.