Few details have been released about the game's storyline other than the fact that the setting will have a similar feel to the movie Black Hawk Down. Takeuchi added that the developing team is composed of staff members who worked on the first Resident Evil. Despite the lack of information, he has stated that Resident Evil 5 will be an actual sequel to the Resident Evil series. Before Takeuchi took over, the game's former producer Hiroyuki Kobayashi stated "it's safe to say that Resident Evil 5 will naturally proceed from where Resident Evil 4 left off."

In a July 2007 Famitsu interview, producer Jun Takeuchi revealed the main playable character as Chris Redfield and the game takes place ten years after the events of the original Resident Evil. The story will explore Chris' life during the ten year interval between the two games. During the game's events, he is a member of a group known as BSAA, and sent to investigate an incident in a desert area that serves as the game's setting. Takeuchi also confirmed that a female character from the series will appear in the game, and that she briefly appears in the full version of the E3 trailer.