Probst was born in Wichita, Kansas, but grew up primarily in Bellevue, Washington. After graduating Newport High School in 1979, he attended Seattle Pacific University, and worked at Boeing Motion Picture/Television studio as a producer and narrator of marketing videos. Probst was divorced from his wife of five years, Shelley Wright, in 2001. In his spare time, Jeff is active in the Elizabeth Glaser Pediatric AIDS Foundation.

In addition to Survivor, Probst previously hosted the FX network's original half-hour show dedicated to answering viewer letters, Backchat (1996). Probst's second television exposure was that of the host of the VH1 series Rock & Roll Jeopardy!, which he hosted for three seasons (from 1998 to 2001), and was a correspondent for the syndicated program Access Hollywood, during which he traveled a reported 300,000 miles. He has also written the screenplay for and directed an independent film, released in 2001, called Finder's Fee. Jeff also does voice work for the animated television program Fillmore, as Vice-Principal Raycliff. People magazine named Probst one of the "50 Most Beautiful People" in 2001. He often contributes to Jeopardy! by giving Survivor related clues from the show's venues, has twice appeared on Celebrity Jeopardy!, first in 2001 and again in 2003, and made several cameo appearances during the April 1, 2010 episode.

He is also a frequent guest star on the sketch show MADtv, guest starring once a season since the show's 9th season.

He has hosted Survivor since 2000. He later stated that he had worked hard to get a meeting with series creator Mark Burnett, where he suggested that Burnett hire him as the host. In this role, he delivers the series' signature catch-phrase to losing contestants, "The tribe has spoken. It's time for you to go."

In 2007, Probst founded The Serpentine Project , a non-profit that empowers youth transitioning out of foster care to reach for their dreams by opening the door to possibilities.

On October 20, 2008, TV Guide reported that Probst is developing a new reality TV series for CBS called Live For The Moment that will feature people with terminal illnesses being taken on "the last adventure of their life" before they die.

On April 1, 2009, Probst appeared on the CBS reality television special I Get That a Lot, in which he worked a cash register.

In February 2010, Probst confirmed that he has signed on to host 2 more seasons of Survivor, putting him through season 22.

In 2005, Probst began dating 26-year-old Survivor: Vanuatu contestant Julie Berry, an East Carolina University graduate who was an athletic scholarship softball player. In a March 2008 interview with the Philippine Daily Inquirer, Jeff revealed that they are no longer together.

Probst was ordained as a minister by the Universal Life Church in 1999. He remarried his parents for their 35th wedding anniversary.