Resident Evil: Revelations marks a return to the survival horror roots of the franchise from the action-centric gameplay of the previous title, Resident Evil 5.

The game takes place before part five in 2005, and after the total destruction of the ultra-modern sea-based city of Terragrigia by terrorists using old Umbrella-style B.O.W.s, or Bio-Organic Weapons. The primary focus of the story is on Jill Valentine and Chris Redfield as they hunt for answers on a mysterious luxury cruise ship known as the Queen Zenobia. The story is broken into chapters fitting for a portable title; story-wise, chapters alternate timelines. Jill Valentine works with her new partner Parker Luciani for the BSAA (Bioterrorism Security Assessment Alliance). Alternate story segments may focus on Chris Redfield and his new partner Jessica Sherawat, or new BSAA members Keith and Quint. Each chapter is intended to gradually wrap together the full narrative.

Exploration in the game is done in third-person 3D, like in Resident Evil 4 and Resident Evil 5, but with the action-heavy gameplay of those titles replaced by more traditional survival horror of the earlier games in the series. Players are once again exploring an ornate and luxurious environment similar to the mansion of the original game, carefully monitoring ammunition, approaching puzzles, and choosing when to fight and when to flee from an attack. The new mutations also appear in smaller numbers, and in claustrophobic areas.

While exploration of the Queen Zenobia is deeply rooted in the survival horror style, many alternating chapters are much more action-oriented and generally resemble the "shooting gallery" segments of Resident Evil 5. These segments may also include much more classic B.O.W. monsters, such as the infamous Hunters and variations of zombie dogs. Ammunition is much more plentiful, and exploration is downplayed.