Transmission experiments were performed with Ixodes scapularis ticks from an uninfected laboratory colony. Immature and adult ticks were exposed to Powassan (POW) viremic hamsters and rabbits, respectively. Oral infection rates for engorged larvae, nymphs and females fed on POW-infected hosts were 10%, 40%, and 57%, respectively. Transstadial transmission rates for nymphs exposed to POW virus as larvae, adults exposed as larvae, and adults exposed as nymphs, were 9.5%, 10%, and 54%, respectively. Evidence of transovarial transmission occurred when two uninfected hamsters, exposed to F2 larvae and nymphs originally exposed to POW virus in the F1 nymphal stage, seroconverted to POW virus with hemagglutination inhibition titers of 80 and 5,120, respectively; the transovarial transmission rate was 16.6%. All developmental stages were able to transmit virus orally to uninfected hosts regardless of when the ticks were originally exposed to the virus. These results suggest that I. scapularis is a competent vector of POW virus under experimental conditions.