North Americans spend half their waking hours engaging in sedentary behaviour. Although several recent interventions suggest that short bouts of uninterrupted sedentary behaviour may result in acute increases in cardiometabolic risk, this literature has not been reviewed systematically. This study performed a systematic review of the impact of uninterrupted sedentary behaviour lasting ≤7 days on markers of cardiometabolic risk (insulin sensitivity, glucose tolerance, and fasting insulin, glucose, and lipid levels) in humans. Interventions were identified through systematic searches of Medline and Embase and screened by 2 independent reviewers. A total of 25 interventions were identified that examined the impact of imposed sedentary behaviour on biomarkers of interest. The majority of these studies focused on healthy young men, with very little identified research on females or other age groups. We found consistent, moderate quality evidence that uninterrupted sedentary behaviour ≤7 days results in moderate and deleterious changes in insulin sensitivity, glucose tolerance, and plasma triglyceride levels. In contrast, there is inconsistent, very low-quality evidence linking uninterrupted sedentary behaviour with changes in insulin, glucose, and HDL- and LDL-cholesterol levels. These findings suggest that uninterrupted bouts of sedentary behaviour should be avoided in order to prevent or attenuate transient increases in metabolic risk.