The film opens in Nazareth where Mary (Gina Rodriguez) is praying. An angel (Joel McCrary) descends from Heaven and tells Mary that she is to bear the son of God. A pygmy mouse named Abby (Kristen Chenoweth) sees the whole thing and spreads the word to the other animals in the town. They look up in the sky and see a bright star.
We meet a young donkey (Steven Yeun) who must work in a wheat mill with his father (Kris Kristofferson). The donkey's dove friend Dave (Keegan-Michael Key) flies in. The two dream of escaping their dull lives and joining the Royal Caravan so that they can ride with the big horses and be adored by crowds, but the Older Donkey scoffs at their dream. The young donkey is confident that he will get out.
Six months later, the donkey is still trapped. When his father realizes how miserable he is and that he has resigned himself to the same fate he has, he loosens the young donkey's collar and knocks out the Miller (Phil Morris) to allow his son to escape. The donkey runs out into the street where Dave catches up with him. The Miller chases after the donkey and nearly catches him, but he manages to hide elsewhere.
Mary is about to celebrate her wedding feast with Joseph (Zachary Levi), but she hasn't told him that she is pregnant. Mary then finds the donkey hiding under a table, believing it to belong to Joseph. He denies that the donkey is his, and Mary notices his leg is injured. After showing him kindness, the donkey takes a liking to her. She decides to name him Boaz, or Bo for short. Joseph then notices Mary's belly, and she comes clean that the baby is meant to be the messiah. Joseph is stunned and confused, but after looking to God for guidance, he promises to help Mary.
Three months later, the Three Wise Men are riding their camels - Felix (Tracy Morgan), Cyrus (Tyler Perry), and Deborah (Oprah Winfrey) - to the home of King Herod (Christopher Plummer). The camels argue over what their trip is for - Felix thinks it's a birthday party, while Cyrus says it's a baby shower, but Deborah says they are going to meet the messiah. The other two think she is crazy. Upon arriving at Herod's home, he thinks the Wise Men's gifts (gold, frankincense, and myrrh) are for him, but one of them says it is for the new king. Feeling threatened, Herod invites (or forces) the men and their camels to stay as guests while he sends his hunter and two dogs - Thaddeus (Ving Rhames) and Rufus (Gabriel Iglesias) - to get rid of this new king.
Thaddeus and Rufus overhear Abby telling the story of Mary to the other mice, and they threaten to eat her until she gives up all the information she knows.
Mary and Joseph leave Nazareth in anticipation of the baby's birth, with Bo and Dave staying behind. The Hunter and the dogs break into their home while Bo tries to hide. Dave tries to create a diversion for Bo to escape, but the dogs spot him. Bo thinks they were sent by the Miller, but he accidentally lets it slip that he was being cared for by Mary. Bo and Dave escape the dogs and run to save Mary.
On their run, Bo stumbles down a cliff. He is helped by a sheep named Ruth (Aidy Bryant), who has abandoned her flock. She joins Bo and Dave as they catch up to Mary and Joseph. Unfortunately, the Hunter and the dogs have found them. Bo pulls Mary toward a market for them to hide. Mary goes by a well to get water, and the Hunter approaches her for the kill. Thinking quickly, Bo pushes a cart down the hill to stop him, but it hits another cart and then causes a chain reaction that destroys most of the market but knocks the Hunter down the well. The villagers are angry at Bo, and Joseph scolds him for what he's done.
Bo decides to go off on his own to try and find the Royal Caravan. Dave follows, and together, they do find it. However, Bo realizes he enjoyed being with Mary and he wants to go back, saying Dave can join them himself. While Dave is displeased at the outcome, he tells Bo that he would have only done it with his best friend. The two head back to help Mary.
Mary and Joseph's cart breaks down, leading them both to confront their fears and doubts over the situation. They resolve to get through it together. Bo returns and picks up all their belongings. Mary then starts to experience contractions and figures that the baby is on his way. Joseph takes all the belongings while Bo carries Mary.
The group arrives in Bethlehem and tries to find shelter, but everywhere is full. The Miller sees Bo and captures him, throwing him in a manger. There, he meets a goat named Zach (Anthony Anderson), a cow named Edith (Patricia Heaton), and a horse named Leah (Kelly Clarkson). They mention not being able to sleep because the light from the star shines brightly through the manger. Bo figures this is where the baby is meant to be born. The three animals help him escape to go get Mary and Joseph to the manger.
Ruth encounters her flock and tries to convince them to help her, but they're still mad at her for abandoning them. The Angel then comes down to tell the shepherds that the messiah is coming, and the sheep suddenly believe Ruth. Meanwhile, Dave encounters the camels after they tried to escape Herod's home, but are now tied to a pole. At the same time, the Hunter and the dogs have found Mary and go to kill her. All the animals come together to stop them and knock them over a cliff. The Hunter hangs on while Bo tries to save Thaddeus and Rufus. The Hunter coldly drops them, but Bo saves them. The Hunter pulls himself up with his sword, but it causes the cliff to break off and send him falling to his death.
The animals go to the manger where Mary has given birth to Jesus. All the animals are in awe. Even Thaddeus and Rufus are so amazed that they resolve to be good dogs. Deborah says this will be a story to be celebrated for thousands of years to come, but Felix and Cyrus still think she's crazy. Mary then invites Bo to go to them and see Jesus. Bo smiles as he rests beside Mary.
*CUT TO THE CHASE*
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The film is a retelling of the Nativity story from the point-of-view of the animals.
Bo the donkey and his dove friend Dave want to escape their dull life in Nazareth by joining the Royal Caravan, but they come across Mary and Joseph as they have just learned that they are to be the parents to the messiah. Meanwhile, King Herod overhears the impending arrival of the new king, and he sends his hunter to kill Mary before she can give birth.
Bo and Dave meet other animal friends as they help bring Mary and Joseph to Bethlehem for the baby's birth. They defeat the hunter and help bring Mary to the manger where she gives birth to Jesus.
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