Anthony Zuiker caught producer Jerry Bruckheimer's attention with his first movie script, convincing him a TV series could be made from the concept; ABC, NBC, and Fox all passed on the pilot before CBS picked it up.

Disney-owned Touchstone Television initially planned to co-produce the show but pulled out once CBS picked it up, since Disney did not want to fund a series for a rival network; Canadian company Alliance Atlantis stepped in as co-producer to save the project.

CBS had a pay-or-play contract with William Petersen, who expressed interest in doing the CSI pilot, which helped convince the network to fast-track the show onto its 2000 schedule.

The show was originally filmed at Rye Canyon, a Lockheed Martin corporate campus in Santa Clarita, California, chosen for its resemblance to the outskirts of Las Vegas, before moving to nearby Santa Clarita Studios after the 11th episode.

After season five, filming shifted primarily to Universal Studios Hollywood, though outdoor scenes continued to be shot around Santa Clarita due to its versatile locations.

The season four episode 'Suckers' was mostly filmed on location in Las Vegas in December 2003, including a Gothic club scene, with additional scenes shot at Caesars Palace in January 2004.

The theme song 'Who Are You' by The Who, sung by Roger Daltrey, led to Daltrey making a special guest appearance in the season-seven episode 'Living Legend' as a character named Mickey Dunn, an episode filled with musical in-jokes including the words 'who's next' on a whiteboard.

A cover of Tears for Fears' 'Mad World' by Michael Andrews and Gary Jules, originally recorded for the film Donnie Darko, was used in the CSI pilot episode and reused in three season-six episodes.

A miniature model prop of Gil Grissom's office, built by the character during season seven, was auctioned on eBay in 2007 and sold for $15,600, with CBS donating the proceeds to a child-advocacy charity.

Fans organized a grassroots campaign in 2007 amid rumors that Jorja Fox might leave the show, raising over $8,000 for stunts targeting CBS executives, including sending 192 chocolate-covered insects and flying a banner plane over Universal Studios; the money was ultimately donated to a child advocacy charity chosen by Fox and producer Carol Mendelsohn.

The so-called 'CSI effect' describes how the show's popularity is believed to have raised jurors' real-world expectations of forensic evidence, with some district attorneys reporting lower conviction rates in cases lacking strong physical evidence.