Sproles was a high school All-American as selected by Student Sports Magazine. He was also the winner of Thomas A. Simone Trophy, given to the top big-class football player in Kansas City metro area. In 1999, he rushed for 2,031 yards in nine games as a junior. In 2000, during his senior year, Sproles rushed for 2,485 yards, scoring 49 touchdowns. He led his Olathe North Eagles to a 12–0 record and their fourth Kansas 6A state title in five years.

Sproles broke over 1000 all-purpose yards in only his seventh game as a Saint. Maintaining an average of 168.5 all purpose yards per game Sproles finished the 2011 season with 2,696 all-purpose yards, breaking the NFL single-season record, previously set in 2000 by Derrick Mason (2,690 yards).

Sproles played college football at Kansas State University from 2001 to 2004, starting all games in his last two seasons and setting 23 school records. In the 2001 season, he made his collegiate debut against New Mexico State and had four carries for 49 yards and a touchdown. He finished with 28 carries for 210 rushing yards and one touchdown in six games.

In the 2002 season, Sproles’s role in the offense expanded. In the season opener against Western Kentucky, he had 135 rushing yards and a rushing touchdown. On October 5, in the game at Colorado, he had 121 rushing yards and three rushing touchdowns. On November 2, against Kansas, he had 110 rushing yards and two rushing touchdowns. He followed that up with 103 rushing yards and two rushing touchdowns against Iowa State. In the next game, against Nebraska, he had 159 rushing yards and three rushing touchdowns. Overall, in the 2002 season, he finished with 1,465 rushing yards and 17 rushing touchdowns.

In the 2003 season, Sproles continued to produce greatly for the Wildcats. In the season opener against California, he had 175 rushing yards and a rushing touchdown. On September 13, against Massachusetts, he had 152 rushing yards and three rushing touchdowns to help get the Wildcats to a 4–0 start. On November 8, in a game at Iowa State, he had 201 rushing yard and three rushing touchdowns. On November 22, against Missouri, he had 43 carries for 273 rushing yards and two rushing touchdowns. In the Big 12 Conference championship in 2003, a 35–7 win over #1 ranked Oklahoma, he had 235 rushing yards and 88 receiving yards and a receiving touchdown. Overall, he finished the 2003 season with 1,986 rushing yards and 16 rushing touchdowns. That same year, he finished fifth in the Heisman Trophy voting. He graduated with a degree in speech pathology. He led all FBS teams in rushing yards in 2003 with 1,986.

Sproles’s father, Larry, played running back at MidAmerica Nazarene University. Larry was 5 ft 5 in (1.65 m) tall. His mother Annette died of cancer in April 2004.

On November 11, 2007, in the Chargers’ 23-21 upset over the Indianapolis Colts, Sproles made history by returning a kickoff and a punt for his first two NFL touchdowns—the first player in league history ever to do so. He returned the opening kickoff 89 yards for the game’s first score. After Nate Kaeding’s field goal increased the Chargers’ lead to 10-0, Sproles returned a Hunter Smith punt 46 yards for another touchdown for a 16-0 lead. Sproles became the first player to return a kickoff and a punt for a touchdown in the same game since Dante Hall did so in 2003.

On December 16, 2007, Sproles ran one yard for his first career rushing touchdown against the Detroit Lions. This was followed by an 11-yard run for his second rushing touchdown, capping off his first career 100+ yard rushing game. Overall, he finished the 2007 regular season with 164 rushing yards and two rushing touchdowns to go along with 10 receptions for 31 yards. On special teams, he had 37 kick returns for 1,008 net yards and a touchdown to go along with 24 punt returns for 229 net yards and a touchdown. The Chargers made the playoffs in 2007. In the Divisional Round against the Indianapolis Colts, he had a 56-yard receiving touchdown in the 28–24 victory. He had 34 rushing yards in the AFC Championship loss to the New England Patriots in the next round.

On September 14, 2008, Sproles became just the second player in NFL history with 50 rushing yards, 50 receiving yards, and 100 return yards in one game (the other being Gale Sayers with the Chicago Bears). His statistics for the game included: 53 rushing yards, 72 receiving yards, and 192 return yards. Sproles logged 317 all-purpose yards in a 39-38 loss to the Denver Broncos. He also had a 103-yard touchdown return. On December 4, he had three receptions for 34 yards and two touchdowns in a 34–7 victory over the Oakland Raiders. In the regular season finale, in the second divisional game against the Denver Broncos, he had 115 rushing yards and his lone rushing touchdown of the regular season. Overall, he finished the 2008 season with 330 rushing yards, one rushing touchdown, and 29 receptions for 342 receiving yards and five receiving touchdowns. On special teams, he had 53 kick returns for 1,376 net yards and a touchdown to go along with 22 punt returns for 249 net yards.

On January 3, 2009, during an AFC Wild Card Round game against the Indianapolis Colts, Sproles ran for two touchdowns in place of the injured LaDainian Tomlinson, including the game-winning touchdown in overtime to beat the Colts 23-17. With 105 yards rushing, 45 yards receiving and 178 return yards, Sproles finished the game with 328 all-purpose yards in the Chargers victory over the Colts, the third most all-purpose yards by a player in a single NFL post-season game in NFL history. He was named the NBC Sunday Night Football “Horse Trailer Player of the Game” for the second consecutive week. The week following however, Sproles was stopped by the Pittsburgh Steelers. He managed only 15 yards on 11 carries, but did have 5 receptions for 91 yards including a 62-yard touchdown.

On February 18, 2009, the Chargers placed their franchise tag on Sproles. He signed the tender on April 28, 2009, which was worth $6.621 million. In Week 2, against the Baltimore Ravens, he had seven receptions for 124 yards in the 31–26 loss. On October 19, he had a 77-yard punt return for a touchdown in a 34–23 loss to the Denver Broncos. On Christmas Day, he had 38 rushing yards and two rushing touchdowns in a 42–17 victory over the Tennessee Titans. Overall, in the 2009 regular season, he finished with 343 rushing yards, three rushing touchdowns, 45 receptions, 497 receiving yards, and four receiving touchdowns. On special teams, he had 54 kick returns for 1,300 net yards and 26 punt returns for 183 net yards and a touchdown.

On March 4, 2010, it was reported that the Chargers would tender Sproles a contract, which was worth approximately $7.3 million. Another team would have had to give the Chargers first- and third-round draft picks if they signed Sproles. He appeared in all 16 games and recorded 59 receptions for 520 yards and two touchdowns to go along with 50 carries for 267 rushing yards. On special teams, he had 51 kick returns for 1,257 net yards and 24 punt returns for 166 net yards.

On July 29, 2011, Sproles signed a four-year contract worth $14 million with the New Orleans Saints. The contract included $6 million in guaranteed money.

On September 8, 2011, during a game against the Green Bay Packers, Sproles returned a punt 72 yards for a touchdown during the second quarter, and returned a second punt for just 20 yards; he also had 2 kick-off returns for 83 yards, including a 57-yard return. That gave Sproles 175 yards of returning on special teams, and on offense, he had 7 receptions for 75 yards, along with just 2 carries for 7 yards, and that gave him 257 all-purpose yards.

On September 25, 2011, during a game against the Houston Texans, Sproles had 193 all-purpose yards (108 special teams yards and 85 offensive yards), with 6 receptions for 50 yards, 2 carries for 35 yards, including a 30-yard touchdown run, and on special teams, had only 1 punt return for 5 yards, but also had 4 kick-off returns for 103 yards.

On October 23, 2011, during a game against the Indianapolis Colts, Darren Sproles had 190 all-purpose yards with 2 touchdowns. On special teams, he had 6 returns for 83 yards (4 punt returns for 27 yards and 2 kick-off returns for 56 yards). On offense, he had 12 carries for 88 yards with a 16-yard touchdown run, and 6 receptions for 19 yards with a 6-yard touchdown pass. During this game, Sproles broke over 1,000 all-purpose yards mark.

On December 26, 2011, during a game against the Atlanta Falcons, Sproles had 236 all-purpose yards. On special teams, he had 4 kick-off returns for 147 yards, including a 92-yard return in which he almost returned the football to the house, and on offense, he had just 89 yards of total offense, with 5 carries for 67 yards, 2 receptions for 22 yards but with a 9-yard touchdown pass from Drew Brees. This last-play touchdown pass in the 4th quarter helped Drew Brees break the single season record for passing yards (5,084), held by Dan Marino since 1984. That pass gave Brees 5,087 yards for the 2011 regular season with one game remaining.

On January 1, 2012, in the last regular season game, a 45–17 blowout of the Carolina Panthers, Sproles had 168 all-purpose yards, in which he had 5 returns for 99 yards (2 kick-off returns for 45 yards and 3 punt returns for 54 yards) on special teams, and broke the NFL record for all-purpose yards in a season (2,696). Sproles played in both of the Saints’ playoff games, scoring 3 touchdowns, including a 44-yard catch and run in the fourth quarter of a celebrated back-and-forth contest against the San Francisco 49ers that the Saints ultimately lost 36-32.

In Week 2, a 35–27 loss to the Carolina Panthers, Sproles had 13 receptions for 128 yards. On November 2, 2012, Saints interim head coach Joe Vitt announced that Sproles would be out indefinitely due to a fractured hand. Sproles returned in Week 12 and played in the Saints’ remaining games, finishing the season with 48 rushes for 244 yards and 1 touchdown, 75 catches for 667 yards and 7 touchdowns, 18 kickoff returns for 483 yards, and 23 punt returns for 183 yards.

In 2013, Sproles played in 15 regular season games, running for 220 yards and 2 touchdowns on 53 carries, making 71 catches for 604 yards and 2 touchdowns, 12 kickoff returns for 225 yards, and 29 punt returns for 194 yards. He played in both of the Saints’ playoff games, with 7 runs for 31 yards and 9 catches for 63 yards.

On March 13, 2014, Sproles was acquired by the Philadelphia Eagles in exchange for a 2014 5th-round draft pick. Drew Brees was quoted after the trade: “You only see a Darren Sproles type of player come around once in your career.”

In 2014, Sproles played in 15 regular season games, running for 329 yards and six touchdowns on 57 carries, making 40 catches for 387 yards, 1 kickoff return for 15 yards, and 39 punt returns for 506 yards and two touchdowns, good for second place on the Eagles’ all-time single-season PR yards, and the most in the NFL in 2014. For his efforts, he was elected to his first career Pro Bowl, where he was drafted as a return specialist but contributed on offense, recording 3 carries for 42 yards and 6 catches for 79 yards. He was also named second team All-Pro as a return specialist, his first time on an All-Pro team.

Starting the 2015 season, much hype surrounded the Eagles signing NFL Rushing Champ DeMarco Murray and San Diego starter Ryan Mathews, leading to speculation that Sproles would get less playing time. In the season opener against the Falcons, Sproles rushed for 50 yards on five attempts and caught seven passes for 76 yards, as well as returning 3 punts for 25 yards. Meanwhile, Murray and Mathews rushed for a combined 11 carries and 13 yards, although they each scored a touchdown rushing while Sproles did not.

On July 29, 2016, Sproles signed a one-year $4.5 million contract extension with the Eagles through 2017. Sproles was an immediate weapon out of the backfield, catching 10 passes for 160 yards and a touchdown in the first three games of the season, however he was also held to only 51 yards on 19 carries rushing in that same timespan. In a Week 8 loss to the Dallas Cowboys, Sproles took over as the lead back, carrying the ball 15 times for 86 yards and catching 5 passes for 17 yards. Sproles would retain the lead back role the following week against the New York Giants, rushing 13 times for 57 yards, catching 3 passes for 14 yards, and gaining 76 yards on 2 punt returns. On November 7, the day after the Giants game, head coach Doug Pederson officially named Sproles as the starting back. Overall, he finished the 2016 season with 438 rushing yards, two rushing touchdowns, 52 receptions, 427 receiving yards, and two receiving touchdowns.

On September 25, 2017, it was revealed that Sproles suffered a broken arm and a torn ACL on the same play in a game against the New York Giants in Week 3. He was officially placed on injured reserve that day, prematurely ending his season. In three games, he rushed 15 times for 61 yards with seven receptions for 73 yards. The Eagles went on to win Super Bowl LII. After the season ended, Sproles stated his interest to return to the Eagles for one more season.

On April 28, 2018, Sproles re-signed with the Eagles on a one-year contract. On July 15, 2018, Sproles announced that 2018 will be his last NFL season. He returned from his injury to play in the season-opening 18–12 victory over the Atlanta Falcons. His return from injury was short-lived as he injured his hamstring in practice before the Week 2 game against the Tampa Bay Buccaneers. He returned in Week 13 to face the Washington Redskins and scored a rushing touchdown. In the following game, against the Dallas Cowboys, he scored a receiving touchdown. He added another receiving touchdown against the Houston Texans in Week 16. Overall, he finished the 2018 season with 120 rushing yards, one rushing touchdown, 160 receiving yards, and two receiving touchdowns. In the Wild Card round against the Chicago Bears, he had 21 rushing yards and 14 receiving yards in the 16-15 victory. In the Divisional Round loss to the New Orleans Saints, he had four rushing yards and 21 receiving yards.

Sproles re-signed with the Eagles on a one-year contract on July 19, 2019. In Week 5, Sproles suffered a partially torn right hip flexor muscle and missed the next three games. He returned in Week 9, but further aggravated the injury. He was later revealed that he fully tore the muscle and was ruled out the rest of the season. He was placed on injured reserve on November 15. On December 21, 2019, Sproles announced that he would retire from the NFL following the 2019 season.

On February 7, 2020, Sproles was hired as a personnel consultant in the Eagles’ football operations department, alongside former teammate Brent Celek.