Van Peebles’ first screen appearance was in 1968, in the soap opera One Life to Live.

In 1971, he appeared in the film Sweet Sweetback’s Baadasssss Song, that Melvin Van Peebles directed and played the lead role in. Mario played his father’s character as a child. The film became a hit and a historical American film, because it was widely credited with showing Hollywood that a viable black audience existed and thus influenced the creation of the Blaxploitation genre. That year, Mario acted in a TV movie called Crosscurrent.

Mario graduated from Saint Thomas More School in Connecticut in 1974 and from Columbia University in 1978 with a bachelor’s degree in economics.

In 1981, Van Peebles acted in the miniseries The Sophisticated Gents.

In 1985, he landed his first leading role in the film Rappin’. He plays Rappin’ John Hood, an ex-convict who attempts to save his neighborhood from developers and hoodlums. That year he also played in the comedy Delivery Boys, was one of the central characters in the action drama South Bronx Heroes, and acted in the dramatic TV film Children of the Night and one episode of the Cosby Show. In 1986, he acted in the urban action film 3:15, the comedy Last Resort, the TV film D.C. Cops, four episodes of L.A. Law, and the Clint Eastwood military film Heartbreak Ridge based on the United States Marine Corps. In 1987, he played in the sport film Hotshot, the TV film The Facts of Life Down Under, and Jaws: The Revenge, the fourth installment of the Jaws franchise.

In 1988, Van Peebles played the lead in the short-lived detective show Sonny Spoon. The show ran for two brief seasons, both of which aired in 1988 before the series was canceled. The show would mark his directorial debut, for which he tackled the task for one episode. That same year, he also acted in the TV film The Child Saver.

In 1989, he directed for the show Top of the Hill, three episodes of 21 Jump Street, and an episode of the TV series Wiseguy.

He made his feature film directorial debut in 1991 with the black gangster film New Jack City, in which he also co-stars. Other lead actors are Wesley Snipes, Ice-T and Judd Nelson. New Jack City was produced with an estimated $8,000,000 budget. The film initially premiered at the Sundance Film Festival on January 17, 1991 before being released nationally on March 8, 1991. It grossed $7,039,622 during its opening weekend. It became the highest grossing independent film of 1991, grossing a total of $47,624,253 domestically. It was also well received by critics. That same year, he directed one episode of Gabriel’s Fire and acted in the TV film A Triumph of the Heart: The Ricky Bell Story.

In 1992, he acted in two TV films: In the Line of Duty: Street War and Stompin’ at the Savoy.

In 1993, he starred in and directed the black Western Posse. It features a large ensemble cast, which includes Woody Strode, Billy Zane, Tiny Lister, Tone Lōc, Big Daddy Kane, Robert Hooks, and many more. The film tells the story of a posse of black soldiers and one ostracized white soldier, who are all betrayed by a corrupt colonel. That same year, he also played in the science fiction crime film Full Eclipse.

In 1994, he collaborated twice with actor Christopher Lambert; they played the side by side leads in the action film Gunmen, and he was the main villain in Highlander III: The Sorcerer, the third installment of the Highlander film series. He also acted in a film called In the Living Years. He followed by directing and co-starring in Panther in 1995. It came from a screenplay adapted by his father, Melvin Van Peebles, from his novel of the same name. The film portrays the Black Panther Party for Self-Defense, tracing the organization from its founding through its decline in a compressed timeframe. Creative license is taken but the general trajectory of the Party and its experiences is factual. At the Locarno International Film Festival it won the prize of the Ecumenical Jury, a Silver Leopard, and was nominated for a Golden Leopard. At the Political Film Society it was nominated for a PFS Award.

In 1996, he acted in one episode of Living Single and one episode of HBO’s Strangers. He also played the lead in the science fiction action film Solo. His directorial release that year was Gang in Blue, in which he stars and co-directs with his father who also has a role in the film. It’s about a black police officer who discovers a cell of white supremacist vigilantes within his department. In 1997, he played in one episode of X-Files. He acted in the films Stag, Riot, and Los Locos, the first American film directed by Jean-Marc Vallée. In 1998, he directed and starred in Love Kills, a comedy about a masseur who gets mixed up in the family plots at the mansion of a recently deceased Beverly Hills millionaire. His acting credits that year were the mini series Mama Flora’s Family, Valentine’s Day, and Killers in the House. In 1999, he acted in Judgement Day, and Raw Nerve.

In 2000, Van Peebles started the new decade by acting in one episodes of Martial Law and 20 episodes of Rude Awakening. In film, he acted in Sally Hemings: An American Scandal and Blowback. In 2001, he starred in a film called Guardian, and he co-starred in the Michael Mann film Ali for which he received a nomination for “Outstanding Supporting Actor” at the NAACP Image Awards. In 2002, he acted in 10,000 Black Men Named George, two episodes of Robbery Homicide Division , and the TV movie Fiona. In 2003, he acted in the film The Hebrew Hammer, 44 Minutes: The North Hollywood Shoot-Out, Gang of Roses, and The Street Lawyer.

His 2003 docudrama Baadasssss!, also known as How to Get the Man’s Foot Outta Your Ass, opened at the Toronto International Film Festival. The film describes the making of his father’s seminal film, Sweet Sweetback’s Baadasssss Song. Mario directed the film as well as portrayed his father in the lead role. The film was critically acclaimed and garnered numerous awards and nominations in the film festival circuits: it won “Best Feature Film” 2004 Philadelphia Film Festival and was nominated for Best Feature 2004 Gijón International Film Festival. At the 2005 Black Reel Awards, he was nominated for Best Actor, won Best Director, won Best Screenplay, and was nominated for Best Film, and the actress Joy Bryant was nominated Best Supporting Actress. Additionally, Van Peebles received three nominations at 2005 Independent Spirit Awards, and two nominations at 2005 NAACP Image Awards.

In 2004, he was a lead in the TV film Crown Heights and acted in one episode of Soul Food. In 2005, he acted in Carlito’s Way: Rise to Power. In 2006, he directed and co-starred in the film Hard Luck, starring Wesley Snipes as the lead, and Cybill Shepherd was also in it. Snipes plays Lucky, a down on his luck former criminal and drug dealer whose post prison trials and tribulations take him on a wild adventure. In 2007, he acted in an episode of Law & Order and the film Sharpshooter. That year, he returned to directing for television starting with three episodes of Damages, in which he also had recurring role until 2009.

In 2008, he acted in his father’s film Confessionsofa Ex-Doofus-ItchyFooted Mutha and 43 episodes of the soap opera All My Children. He returned to Law & Order as a director for three episodes. In 2009, Van Peebles and his family starred in the reality TV show Mario’s Green House. He also acted in the film A Letter to Dad.

In 2010, he acted in two films Multiple Sarcasms and Across the Line: The Exodus of Charlie Wright. He directed one episode of Lost, the documentary Fair Game?, and the film Redemption Road. In 2011, he directed the sport drama All Things Fall Apart starring rapper/actor 50 cent in the title role playing a football player who suffers from a deadly disease; Van Peebles also played a role in this feature. His acting credits that year were the films Tied to a Chair and 5th & Alameda. He also acted in two episodes of Hellcats and two episodes of The Game. That year, he also started to direct for the TV series Boss and would direct five episodes until 2012.

In 2012, he wrote, directed, and acted in “We the Party”, a comedy set in an ethnically diverse Los Angeles high school. The film focuses on five friends as they deal with romance, money, prom, college, sex, bullies, Facebook, fitting in, standing out, and finding themselves. Also that year, he acted in one episode of The Finder and the feature film American Warship. In 2013, he directed one episode NCIS: Naval Criminal Investigative Service, one episode of Monday Mornings, and one episode of Zero Hour. Also that year, he directed an episode of Nashville. The following year, he directed two additional episodes of Nashville the following year as well acted in one, and in 2015, he reprised his role with another director.

In 2014, Van Peebles wrote, directed, and acted in Red Sky. The film is an action/thriller starring Cam Gigandet, Shane West, and Rachael Leigh Cook. He also directed three episodes of Once Upon a Time, and that year, he acted in two other films Mantervention and Drumline: A New Beat.

In 2015, he directed one episode of The Last Ship, one episode of Chicago P.D., and two episodes of Empire. In 2016, he directed another episode of Empire and two episodes of Being Mary Jane. In 2017, he directed two additional episodes of Being Mary Jane and one episode of Hand of God. He also acted in the feature For Justice. Also in 2016, he directed the film USS Indianapolis: Men of Courage, based largely on the true story of the loss of the ship in the closing stages of the Second World War and starring Nicolas Cage, Tom Sizemore, and Thomas Jane. He directed one episode of Roots and acted in the feature film Submerged.

In 2017, he acted in four episodes of Bloodline, one of which he also directed, and he directed an additional episode his character didn’t appear in. That year, he created and produced the TV series Superstition, directed eight episodes, and acted in ten, and in some of them he performed both roles.