Bo Peep is a sweet and lovable porcelain shepherdess figurine. Bo Peep and her sheep are adornments of Molly's bedside lamp. She is inspired by the character of the classic children's nursery rhyme, Little Bo-Peep. Bo is sometimes considered to be "Andy's toy", because Andy likes to make her the damsel in distress in his plays. Bo is the romantic interest (later girlfriend) of Woody, providing a calm and loving comfort whenever he is overwhelmed. She is noted for using her shepherdess crook to hook her cowboy and bring him closer in a romantic way. In private, she is a great deal more daring and seductive with her words and actions, which Woody seems to be quite fond of. She seems to have faith in Woody, which is shown when she assures him that Andy, who is excited to have Buzz Lightyear, will still have a special place for him and that whether if Woody has his hat or not, Andy will still take him out to Cowboy Camp. In Toy Story, she is one of the very few toys in Andy's room who believed that Woody would not intentionally harm Buzz when he accidentally pushed him out of the window. She is shown to be shocked upon hearing about Woody's disappearance when Andy comes back home from Pizza Planet. On the night before the moving day, Bo Peep is shown to be worried about Woody when she sees a sad Andy sleeping. Later, after witnessing Woody being thrown out of the moving truck under Mr. Potato Head's orders, Bo Peep helps the toys realize that "Woody was telling the truth," and the toys brighten up. In the end of Toy Story Bo Peep and Woody kiss under a mistletoe. In Toy Story 2, she is shown feeling devastated after Al McWhiggin of Al's Toy Barn steals Woody. When Buzz plans a rescue, Bo Peep gives Buzz a kiss (intended for Woody when Buzz finds him) and waves goodbye to Buzz and the other toys, wishing them good luck, as they leave on their mission to rescue Woody. After the toys return home, Woody shows her his arm fixed by Andy himself, which she greatly admires. Finally, she and Woody are seen together with Buzz and Jessie as they enjoy watching Wheezy sing "You've Got a Friend in Me."

In Toy Story 3, it is revealed that Bo Peep, along with many of Andy's other toys, were either sold, missing, broken or thrown away years after Toy Story 2 so she only appears via archive footage in Toy Story 3.