Peter Rabbit is an 8 year old rabbit who lives with his mother underneath a tree in a hidden burrow. He misses his father and wants to grow up to be just like him. Armed with his father's journal, which is basically a guide to everything one needs to become a truly wild rabbit, and aided and abetted by his two best friends, Benjamin Bunny and Lily Bobtail. Peter sets off to make his own mark in life and have adventures even greater and grander than his dad's.
- Peter Rabbit - A mischievous, adventurous, quick witted and street smart young rabbit who wants to be just like his father and goes on his adventures with his friends. Older cousin of Benjamin. He is also Lily's potential love interest. (Voiced by Colin De Paula (Season 1), L. Parker Lucas (Season 2) )in the USA and Connor Fitzgerald in the UK).
- Benjamin Bunny - A loyal, kind hearted rabbit, who is Peter's younger cousin and friend who joins him on his adventures. He often gets extremely scared, worried and afraid of danger, and he's a bit clumsy and carefree. (Voiced by Peter Harris in the USA and Danny Price in the UK).
- Lily Bobtail - A quick-witted and book smart female rabbit who always has an idea or item to any problem, she is also best friends with Peter and Benjamin. She is also Peter's potential love interest. (Voiced by Michaela Dean in the USA and Harriet Perring in the UK).
- Mr. Tod - A clever, sneaky fox who is always trying to capture Peter Rabbit and his friends. (Voiced by Mark Huckerby).
- Squirrel Nutkin - A mischievous, tailless squirrel who is the leader of the squirrels, he is also friends with Peter, Benjamin and Lily and will often help them whenever their in trouble. (Voiced by Kyle D. Massey and Nigel Pilkington).
- Mrs. Josephine Rabbit - Peter's kind-hearted loving mother who supports and often gives advise to him about any adventures he's on. (Voiced by Stephanie Sheh (Season 1) and Katie Koslowski (Season 2)).
- Flopsy Rabbit and Mopsy Rabbit - Two twin sisters of Peter Rabbit and are always with each other. (Voiced by Megan Harvey (Season 1) and Brittany Harms and Sawyer Niehaus (Season 2).
- Cotton-Tail Rabbit - A very young rabbit who is the baby sister of Peter, Flopsy and Mopsy, she looks up to her big brother and always wants to go on adventures with him. (Voiced by Jenna Iacono (Season 1) and Aviella Kibel (Season 2).
- Mr. Jeremy Fisher - (Voiced by Kirk Thornton).
- Jemima Puddle-Duck - (Voiced by Sarah Bolt).
- Mrs. Tiggy-Winkle - (Voiced by Gwenfair Vaughan).
- Ginger And Pickles - (Voiced by J.B. Blanc and Kirk Thornton).
- Mr. McGregor - (Voiced by Dave Mitchell).
- Tommy Brock - (Voiced by John White).
- Samuel Whiskers - (Voiced by Shawn Curran).
- Old Brown - (Voiced by Dave Mitchell).
- Princess Hazel Rabbit - (Voiced by Carly McKillip)
- Dr. Bobtail -
- Mrs. Bobtail -
- Lily Bobtail is a new character created for the show, she didn't appear in the books like Peter and Benjamin did.
- Unlike the books, Peter Rabbit doesn't wear shoes.
- Mrs. McGregor hasn't been mentioned on the show yet, so it's unknown if Mr. McGregor is married or not.
- Why Peter Rabbit's father isn't on the show hasn't been mentioned yet, but it's assumed that he's dead.