Rolle was born in Homestead, Florida. He attended South Dade High School in Homestead, where he was a letterman in football and played for the South Dade Buccaneers high school football team. As a junior, he posted three interceptions, 85 tackles, and five pass deflections, and he returned three kickoffs and three punts for touchdowns. As a senior, he compiled three interceptions and 86 tackles; on offense, he had four receptions for 160 yards and a touchdown. In track & field, Rolle was a state qualifier in the hurdling events. At the 2000 FHSAA Outdoor State Finals, he took 10th in the 110-meter hurdles (15.46 s) and 15th in the 300-meter hurdles (41.84 s).
Rolle attended the University of Miami, where he played for coach Larry Coker's Miami Hurricanes football from 2001 to 2004. During his 2001 freshman season, Rolle was one of four true freshman to letter at Miami. He appeared in eight games, recording eight tackles and an interception. Rolle started eleven games as a sophomore in 2002, earning first-team All-Big East honors. Rolle totaled a career-high 66 tackles with two sacks, six tackles for a loss, seven pass deflections, and an interception for the season.
In 2003, as a junior, Rolle recorded 51 tackles, 3.5 tackles for a loss, seven broken up passes, and two interceptions on the year. In his final season, 2004, Rolle was recognized as a unanimous All-American. He recorded 58 tackles his senior year, 6.5 tackles behind the line of scrimmage, six pass deflections, and an interception. He was hurt late in the '04 season and missed the Virginia Tech game because of a foot sprain.
Following his senior season in 2004, he was a first-team All-Atlantic Coast Conference (ACC) and a consensus first-team All-American at cornerback. His notable senior performances included shutting down talented opposing receivers such as Larry Fitzgerald (three catches for 26 yards) and Calvin Johnson (two catches for 10 yards); both were future All-Americans.
The Arizona Cardinals selected Rolle in the first round (eighth pick overall) of the 2005 NFL Draft, making him the second cornerback picked. Rolle's career did not start off well mainly because of torn cartilage in his left knee suffered September 25, 2005 in a game against the Seattle Seahawks. The injury required surgery on October 3, 2005 and put Rolle out for almost the entire 2005 NFL season.
Rolle returned and played on December 11, 2005 against the Washington Redskins, where he recorded his first interception and returned it for 29 yards. He played in week 14 and again in week 15 against the Houston Texans.
The 2007 campaign did not start well for Rolle as he lost his starting spot in the preseason. However, he battled back and had the highlight of his career on November 18 when he intercepted 3 passes by Carson Palmer of the Cincinnati Bengals, returning the first two for touchdowns of 55 and 54 yards. He returned the third interception all the way to the end zone as well, but it was called back on a penalty. Rolle was rewarded for his performance when he was named NFC Defensive Player of the Week. Rolle finished the season with 61 tackles, 5 interceptions, and three return touchdowns. His interception total was tied for ninth most in the NFL. Cardinals Head Coach Ken Whisenhunt announced on February 2008 that Rolle would be moved to free safety for the 2008 season. For several plays during the 2009 season, Rolle was used as the quarterback in a Wildcat offense.
On March 4, 2010, the Cardinals released Rolle in order to avoid a $4 million roster bonus.
On March 5, 2010, the New York Giants signed Rolle to a five-year, $37 million contract. This deal made Rolle one of the highest paid safeties in the NFL.
In the 2011 regular season, Rolle compiled a career-high 96 tackles. Antrel Rolle was also part of the New York Giants Super Bowl team that won against the New England Patriots in Super Bowl 46, 21-17. In 2012, Rolle was named the NFC Defensive Player of Week 6.
In 2013, Rolle led the league's safeties in interceptions with six and was named to the Pro Bowl.