Formation of a single crystalline oxide semiconductor on an insulating film as a channel material capable of three-dimensional (3D) stacking would enable 3D very-large-scale integration circuits. This study presents a technique for forming single-crystalline In2O3 having no grain boundaries in a channel formation region on an insulating film using the (001) plane of c-axis-aligned crystalline indium gallium zinc oxide as a seed. Vertical field-effect transistors using the single-crystalline In2O3 had an off-state current of 10−21 A μm−1 and electrical characteristics were improved compared with those using non-single-crystalline In2O3: the subthreshold slope was improved from 95.7 to 86.7 mV dec.−1, the threshold voltage showing normally-off characteristics (0.10 V) was obtained, the threshold voltage standard deviation was improved from 0.11 to 0.05 V, the on-state current was improved from 22.5 to 28.8 μA, and a 17-digit on/off ratio was obtained at 27 °C.