Nanomaterials have extensive applications in the development of sensitive biosensors, but the influence of their specific structural properties remains unclear. This work presents a platform that can provide mechanistic insight into how nanostructured electrodes improve the performance of electrochemical biosensors. We designed nanoelectrodes with sub-10 nm spike features through a combination of top-down lithography and solution-based synthesis. These anisotropic structures facilitated rapid electron-transfer, minimized biofouling, and promoted efficient target capture. Using these spiky nanoelectrodes in a biosensor, we detected bacterial mRNA at aM-levels and within 3 min. Our findings reveal the mechanism underlying signal enhancement from high-curvature regions on nanostructured electrodes, highlighting the structure–property relationships of nanostructures in electrochemical sensing.