Colloidal quantum dots are sub-10 nm semiconductors treated with liquid processes, rendering them attractive candidates for single-electron transistors operating at high temperatures. However, there have been few reports on single-electron transistors using colloidal quantum dots due to the difficulty in fabrication. In this work, we fabricated single-electron transistors using single oleic acid-capped PbS quantum dot coupled to nanogap metal electrodes and measured single-electron tunneling. We observed dot size-dependent carrier transport, orbital-dependent electron charging energy and conductance, electric field modulation of the electron confinement potential, and the Kondo effect, which provide nanoscopic insights into carrier transport through single colloidal quantum dots. Moreover, the large charging energy in small quantum dots enables single-electron transistor operation even at room temperature. These findings, as well as the commercial availability and high stability, make PbS quantum dots promising for the development of quantum information and optoelectronic devices, particularly room-temperature single-electron transistors with excellent optical properties.