Place it at the end of a short rejection scene, right after the request appears on screen, then follow it with No No Wait Wait for a second refusal and Knuckles Oh No when the idea gets worse. That order builds the joke in three steps without repeating the same reaction.
About this sound
This is a short vocal reaction built around a flat 'No' that slides into a long, wobbly 'Creooo', delivered like a cartoonish groan of refusal. It feels playful and over-the-top, turning a simple rejection into a joke that lands because the sound drags on just a little too long.