The Giants, who beat NFC East rival Dallas 31-14 in their last regular season game to win the division and qualify for the play-offs, will host the Cowboys at MetLife Stadium on September 5.
It will mark the ninth consecutive season that the league's defending champion hosts the NFL Kick-off game, with the home team winning each of the previous meetings.
Unlike previous years when the NFL opened its season on a Thursday, the Giants-Cowboys game is being played on a Wednesday to avoid a conflict with US President Barack Obama's Thursday night speech at the Democratic National Convention.
The Giants finished the 2011 regular season with a 9-7 record before sweeping four post-season games to win their second championship in five seasons.
Dallas, who would have advanced to the play-offs with a victory over the Giants in the season finale, finished 8-8.