In the Ohio River Valley, War Chief Pontiac and a large alliance attacked numerous British forts and settlements as a reaction to the offensive policies of British General Jeffrey Amherst. Hostilities came to an end after British Army expeditions in 1764 led to peace negotiations over the next two years. Native Americans were unable to drive away the British, but the uprising prompted the British government to modify the policies that had provoked the conflict.

Warfare on the North American frontier was brutal, and the killing of prisoners, the targeting of civilians, and other atrocities were widespread. In what some historians consider an incident of biological warfare, British officers at Fort Pitt gave smallpox infested blankets to the besieging Native Americans with the intent of spreading the disease, possibly contributing to the smallpox epidemic that killed much of their population.

The ruthlessness and treachery of the conflict was a reflection of a growing divide between the separate populations of the British colonists and Native Americans. Contrary to popular belief, the British government did not issue the Royal Proclamation of 1763 in reaction to Pontiac's War, though the conflict did provide an impetus for the application of the Proclamation's Native clauses. This proved unpopular with British colonists, and may have been one of the early contributing factors to the American Revolution.