In November 1993, Rodriguez appeared in one episode of the daytime drama All My Children. In August 2005, he made a two-day guest appearance on the ABC daytime drama One Life to Live.

As a singer, Rodriguez has performed on stage and as a recording Artist. In 2002, he created his own musical cabaret show, titled Monday Night Twisted Cabaret, which ran at New York gay club xl for a year. Rodriguez continued to perform Twisted Cabaret on a sporadic basis between 2004–2006.

In 2003, Rodriguez became Queer Eye's resident "Culture Vulture", functioning as the show's expert on the topic of culture, and the show's success brought him increased exposure. He was a replacement host for the show's originally-cast culture host, but has been part of Queer Eye since the third episode. This show includes five guys, known as the "Fab Five." The roles of the "Fab Five" are a grooming guru, food and wine connoisseur, fashion savant, a designer Doctor, and a culture vulture. The purpose of the show is to create a better sense of fashion, function, and culture between all straight men. A very interesting aspect of the show is the chemistry and great casting of the "Fab Five." These actors seem like they have been the best of friends since drama camp began back in time. However, none of the men knew each other before the show started. Since Jai Rodriguez is the youngest guy in the clique by ten years, he is considered to be the unique one. He came into the show after it had already begun. Also, Jai is the only person of color on the show.

Rodriguez is also a co-author of the New York Times bestselling book, Queer Eye for the Straight Guy: The Fab Five's Guide to Looking Better, Cooking Better, Dressing Better, Behaving Better, and Living Better (Published by Clarkson Potter, 2004).

In November 2005, while Queer Eye was on filming hiatus, he joined the cast of The Producers on Broadway for three months, playing the role of Carmen Ghia. He played a different role, that of Sabu, in the 2005 Producers movie.

Rodriguez appeared as one of the Celebrity Singers on the Fox reality TV show Celebrity Duets, which premiered August 29. He was eliminated on September 22, 2006.

Rodriguez released his first single, "Love Is Good", on the indie record label Arrive at Eleven Productions in March 2003. He then recorded his debut album for Airgo Records. The first single from the album, Broken, was released on October 4, 2007.

Rodriguez also hosted the Animal Planet reality TV show Groomer Has It, which premiered April 12, 2008 on Animal Planet. He also appears on GSN Live as a sub host.

In March 2010, Rodriguez appeared as a newscaster in the music video for "Telephone", with Lady Gaga and Beyoncé.

In November 2011, Rodriguez appeared in an episode of How I Met Your Mother called "The Rebound Girl", playing the husband of Barney Stinson's brother. He appeared again in March 2014 in the episode "Gary Blauman".

As of May 2012, the new ABC sitcom Malibu Country has been picked up. Rodriguez is a regular cast member, playing record industry executive, Geoffrey.

From August 2014, Rodriguez appears as "Ritchie Valentine" in The Horizon.