Al Kapone's involvement in the film came from a case of mistaken identity. Director Craig Brewer was expecting a phone call from DJ Paul (Paul Beauregard) because he wanted to hire Paul to write the songs that DJay would perform. Kapone, who knew Brewer from the Memphis music scene, decided to call him at that very same time. He told Brewer that the movie needed to have his music in it and Brewer immediately agreed. After a few minutes of small talk, Brewer realized he was talking to the wrong person. Too embarrassed to back out of the deal, Brewer told Kapone that he could audition with one song. Kapone had only 24 hours to write a song for DJay. He was sent the script by courier and was given Terrence Howard's phone number to discuss the character. The next day, Kapone performed "Hustle & Flow (It Ain't Over)" for Brewer and producer John Singleton. They loved the song so much, they used an additional three of Kapone's songs for the soundtrack.
While filming in Memphis, Tennessee, Anthony Anderson and assistant director Wayne Witherspoon were arrested and charged with sexual assault on a woman who Witherspoon allegedly lured to a production trailer. Charges against Anderson were later thrown out by the presiding judge.
There are numerous references to Memphis-based musicians in the film: The character of Shelby was named after Shelby Bryant, a singer-songwriter from Memphis. In one scene, Shelby is wearing a T-shirt with the logo of Sam Phillips's Memphis Recording Service studio. In another scene, Shelby wears a T-shirt of Lucero, a local country-rock band. Many of the studio musicians who played for Stax Records, the legendary Memphis-based record label, play on the original score. Isaac Hayes, who plays Arnel, recorded for Stax Records. The cover of the 1974 Stax album "Victim of the Joke?" by Memphian David Porter is stapled to DJay's work table. Otis Redding, who also recorded for Stax Records, is mentioned in one scene. Al Green's song "Jesus is Waiting" is heard during one scene. Memphis native Josey Scott of the band Saliva, appears as a store owner. Members of the popular rap group Three 6 Mafia appear in the film. Paul Beauregard plays DJay's neighbor and Jordan Houston plays Skinny Black's brother. Haystak, Free Sol and Al Kapone are local rappers who appear in the film.
Although there are numerous references to famous Memphis-based musicians in the film, Craig Brewer deliberately avoided any direct references to Elvis Presley. In an interview, Brewer said: "That was a rule. No Elvis."
Terrence Howard interviewed 123 pimps and 78 prostitutes over a period of two and a half years. This process included living with four separate pimps for various periods, including a month-long stint in a Memphis bordello.
Craig Brewer added several touches from his personal life in the script: his wife worked in a strip club, then got pregnant, he would have to turn off the air conditioning to edit or the fuse would blow out and he actually saw a black pimp with a white braids-wearing hooker in a car trying to hustle up some business near a local hotel.