The study determined the risk zone and estimated the population at risk of adverse health effects for arsenic exposure along the bank of River Indus in Pakistan. A cross-sectional survey was conducted in 216 randomly selected villages of one of the districts along River Indus. Wells of ten households from each village were selected to measure arsenic levels. The location of wells was identified using global positioning system device, and spatial variations of the groundwater contamination were assessed using geographical information system tools. Using layers of contaminated drinking water wells according to arsenic levels and population with major landmarks, a risk zone and estimated population at risk were determined, which were exposed to arsenic level ≥10 µg/L. Drinking wells with arsenic levels of ≥10 µg/L were concentrated within 18 km near the river bank. Based on these estimates, a total of 13 million people were exposed to ≥10 µg/L arsenic concentration along the course of River Indus traversing through 27 districts in Pakistan. This information would help the researchers in designing health effect studies on arsenic and policy makers in allocating resources for designing focused interventions for arsenic mitigation in Pakistan. The study methods have implication on similar populations which are affected along rivers due to arsenic contamination.