The game is separated into two sides: the U.S.S. side and the SPECOPS side (DLC). Each side has six chapters, where players can choose four of the six playable characters per side.

U.S.S. Delta team (also known as "Wolfpack") enters the Raccoon City Underground Laboratory, where they meet up with Alpha team leader HUNK. Their mission to assist the Alphas in stopping Dr. Birkin from handing over his G-Virus research to the US Military and retrieve the G-Virus. On their way to Birkin's lab, they find that Birkin has paid-off numerous UBCS mercenaries into working for him while the deal goes forward. When they reach Birkin's lab, the doctor is fired upon from behind a closed door and HUNK and another Alpha leave with the samples. Heading up the elevator to meet up with Goblin 6, they soon find that Birkin has survived the attack and infected himself with the virus. The creature proceeds to kill off most of Alpha team before disappearing; HUNK offers to go back in search of the sample.

Not long after the battle, it becomes evident that the t-Virus has leaked city-wide, and people begin to transform into zombies soon after. In what they see as a punishment by USS Command, Delta team is ordered to remove evidence of Umbrella's role in the outbreak. Heading into Raccoon City Hall, Delta Team meets UBCS mercenary and monitor Nicholai Zinoviev; he is soon revealed as a traitor and attempts to kill the Wolfpack.

Later, the team is sent out around Raccoon Park to find the Nemesis-T Type, which has gone rogue. A second parasite is injected into its body in order to bring it back under control. Shortly after this mission is completed, the team is then sent out to the Raccoon City Police Department, ordered to kill any surviving police officers and destroy evidence linking the company to the outbreak. When this is all done, the team exits the station, and soon after encounter Leon S. Kennedy, whom they begin to hunt down

Wolfpack

An elite team of the Umbrella Security Service sent to kill any survivors and destroy all the evidence about the company's culpability for the infamous outbreak.

"VECTOR" - Master of stealth, real name unknown
Hector Hivers - Explosives expert, codename "BELTWAY"
Karen LesProux - Team leader, codename "LUPO"
Vladimir Bodrovski - Designated marksman, codename "SPECTRE"
Michaela Heinzwaffen - Team medic, codename "BERTHA"
Christine Yamata - Field scientist, codename "FOUR EYES"

Echo Six

An elite squad of the SPEC OPS FIELD UNIT sent by the US Government to find the truth about the outbreak and rescue the civilians of Raccoon City.

Caroline Floyd - Recon, codename "Willow"
Crispin Jettingham - Team leader / Assault, codename "Dee-Ay"
Marissa Ronson - Demolitions expert, codename "Tweed"
Erez Morris - Team medic, codename "Harley"
Lawrence Kimbala - Field scientist, codename "Shona"
Sienna Fowler - Marksman, codename "Party Girl"

Classic characters

Main article: Heroes Mode

Currently, seven playable classic characters from Resident Evil 2 and Resident Evil 3 were confirmed as playable in the new Heroes Mode and encounters in the main story line of the game. Characters like Leon, Claire and Jill can be killed by the USS to change the events of the series, meanwhile HUNK, Ada and Nicholai serves as support in specific chapters.

Also, a new character nicknamed LONE WOLF was confirmed to enter in the Heroes mode.

Leon S. Kennedy - A rookie cop and one of the surviving Raccoon City Police Department, and as a cop, he is targeted by the U.S.S.
Claire Redfield - College student and sister to Chris Redfield. She was "prosecuted" by the WOLFPACK.
Jill Valentine - A former S.T.A.R.S. Alpha Team member and a survivor of the "mansion incident".
Carlos Oliveira - One of the few surviving U.B.C.S. mercenaries, he was confirmed in the same trailer as Jill.
HUNK - A U.S.S. veteran member known as "Mr. Death", confirmed via Inserted Evil clue #4; the Wolfpack met with him after he obtained a G-Virus sample.
Nicholai Zinoviev - A Russian mercenary of the U.B.C.S., he was confirmed to play a important role in the main game.
LONE WOLF - A legendary helicopter pilot of Umbrella, it is unknown if he appears in the main game, but he was confirmed as a playable character in the Heroes Mode.
Ada Wong - A female spy sent by The Organization to obtain a G-virus sample.