Morrison was born in Fort Ord, California. The inspiration for his career as an actor came from attending theatre camp as a child and acting in school plays and musicals. He was also a soccer/football player and was elected prom king and president of his high school. He also dated the school's prom queen.

He was educated at New York University's Tisch School of the Arts and the Orange County High School of the Arts, while at the Los Alamitos High School campus.

Morrison's career began when he made his musical theatre debut on Broadway in the musical version of Footloose, followed by a revival of The Rocky Horror Show. However, his big break came when he landed the role of Link Larkin in the Broadway production of John Waters' Hairspray. After performing in the role for some time, he started working in television, guest-starring on shows such as Ghost Whisperer, Numb3rs, CSI: Miami and Hack. He also had small roles in films such as Marci X, Primary Colors, and Simply Funk. He followed up by performing the role of Sir Harry in the ABC-TV production of Once Upon a Mattress, starring Tracey Ullman and Carol Burnett.

In 2001 Matthew Morrison was recruited to complete the quartet for the band LMNT. He was replaced instead by Jonas Persch by the time their first album was released. He commented in an interview about his experience in the boy band by remarking, "It was the worst year of my life. You know when you're a performer and you're out there on stage and you're embarrassed that you're doing something wrong. It was pretty bad."

In 2002, Morrison starred on the Original Cast of the musical Hairspray as Link Larkin. He would remain in this role until January 2004.

In 2005 Morrison changed gears, playing the role of Fabrizio Nacarelli in Adam Guettel's The Light in the Piazza. He was nominated for a Tony Award for his performance. He left the show August 28, 2005.

Morrison joined the cast of the CBS soap opera As the World Turns as Adam Munson on October 24, 2006, but left abruptly soon after due to a "scheduling conflict," last airing on the soap on November 27.

Morrison appeared in a benefit performance celebrating Andrew Lippa, and the 25th Anniversary of the performing troupe 'The Kids in the Hall' at the Orange County Performing Arts Center in Costa Mesa, California in December 2006. The performance benefited The California Conservatory of the Arts, a non-profit arts education organization focusing on pre-professional training for young students, grades six through twelve, in Orange County, California.

He earned a Drama Desk Nomination for his role in the Off Broadway show 10 Million Miles which ran at the Atlantic Theater Company in 2007. That same year he appeared in two films: Dan in Real Life, followed by Music and Lyrics as Cora Corman's manager.

Morrison was engaged to actress Chrishell Stause.