The purpose of this study was to quantify running economy (RE) during level treadmill running in 6-year-old children and to identify the potentially mediating effects of resting oxygen uptake and body fat percentage on sex differences in RE. Resting oxygen uptake (VO2), body fat, and RE at 5 mph were quantified in 15 boys and 20 girls following 30 min of treadmill accommodation. While absolute VO2 and mass-related values of gross and net VO2 were significantly higher in boys compared to girls, gross VO2 expressed relative to fat-free mass was not different between sexes. These results indicate that 6-year-old girls exhibit better RE compared to 6-year-old boys when VO2 is expressed as a function of total body mass. This sex difference in VO2 may reflect an increase in aerobic energy demands associated with the presence of a greater muscle mass in boys.