A government statement published on Monday announced that Vidal's position will be taken by the "experienced diplomat" Carlos Fernandez de Cossio.
"Fernandez de Cossio is one of the most complete Cuban diplomats," said Johana Tablada, the deputy head of US relations, on Twitter.
A former ambassador to Canada and South Africa, Fernandez de Cossio already held this position at the US office during the 1990s, at a time of great tension between Washington and Havana.
According to the statement, Vidal handed over her duties to Fernandez de Cossio on Feb. 9 after a formal ceremony.
"In her almost 12 years at the Directorate-General of the US Office, Josefina Vidal carried out her complex job with efficiency, talent and sensibility," read the statement.
Josefina Vidal was considered the Cuban face of the long process of negotiations that concluded in the restoration of US-Cuba diplomatic relations in August 2015.
After 54 years of political enmity, respective embassies in Havana and Washington were re-opened, and former president Barack Obama visited the Caribbean nation in March 2016.
However, relations have soured once again since President Donald Trump promised to roll back formal ties "in search of a better deal with Havana."
Thus, Fernandez de Cossio returns to his previous office in similar conditions to those he had to deal with two decades ago.
The statement did not clarify what functions Vidal will be taking on.