The Knife is a melee weapon in Resident Evil 5. Several of the context sensitive "Help" animations depict the characters using their knives to rescue their partners from bad situations, such as being absorbed into Uroboros, impaled by a Licker β's tongue, or latched on to by Bui Kichwa. Each character uses their own knife and they all do the same amount of damage to enemies. Sheva is also the only character who uses the knife in some of her various melee attacks, Impale, Throat Slit, and Double Fang.

Compared to its RE4 counterpart, the knife does very little damage - it's a bit slower than the knife from the previous game and it only deals 50 damage per strike, similar to an un-upgraded VZ61, . The Majini's are also heavily armed with large kukri knives, and axes.

The award "Be The Knife", (Achievement for Xbox 360 and Trophy for PS3) can be earned by hitting away a Bowgun arrow with your knife. There's another achievement/trophy called "A Cut Above" and it's awarded when the player kills five enemies with the knife.

Jill Valentine does a jab attack with her knife instead of a slash like other characters do, and attacks considerably faster with it. Rebecca Chambers swings her knife slightly slower than other characters.

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