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The Pranksters are the Electric Company's enemies. In the Prankster Cam segments, each of the Pranksters explain about the letters, with Manny specializing in punctuation and Annie talking about apostrophe-S. They are also most likely to be vandals, especially towards the Electric Company, and some others' property. Despite all their efforts, they are always defeated in the end by the Electric Company.
Even though they try to pull pranks on the Electric Company, on very rare occasions, The Pranksters will sometimes provide support. Some examples include Manny Spamboni who helped the Electric Company fix an incubating system to hatch an egg.
- Francine Carruthers (played by Ashley Austin Morris) - The vandalistic leader of the Pranksters. She has the same power as the Electric Company—the ability to generate word balls, except that hers are violet. Francine has a very high opinion of her own intelligence. Most of her plots involve making herself look good at the expense of the Company. In Season 3, she gets an assistant named Gilda Flip. She usually calls Marcus "Marty Farms" and usually gets 100 to 200 presents for her birthday.
- Annie Scrambler (played by Sandie Rosa) - A girl who is frequently envious of the accomplishments of others. She has the power to scramble any word or sentence with a stomp of her foot which results in making new words, but sometimes she scrambles existing words to create nonexistent ones. Annie is often jealous of the Electric Company's good luck and sometimes works with her Uncle Sigmund, who is a noted hypnotist, to get back at her rivals. She will work with the Electric Company if she has to, such as to help her uncle out in "Bananas". Annie is portrayed by Sandie Rosa.
- Danny Rebus (played by William Jackson Harper) - A suave, sophisticated Prankster. He can turn any sentence into a rebus puzzle. Most of his messages make fun of the Electric Company. Danny is very proud, yet has a touchy personality, being easily offended. He often accuses the Electric Company of making him look bad, promising vengeance. He has helped the Electric Company out on occasion when it benefits him. He is also the most destructive member.
- Manny Spamboni (played by Dominic Colón) - The loudest of the Pranksters. He is the only member of the group that does not have any powers, but he is quite skilled in the art of robotics and mechanics. Manny is rude and crude, and uses his many gadgets to make trouble for the Electric Company. He is thought the world of by his mother who says he is her "little angel," although she has put him in his place on occasion, such as chastising him for cheating at a limerick competition.
- Gilda Flip (played by Carly Rose Sonenclar) - The newest member of the Pranksters and Francine's assistant. Like Manny, she doesn't appear to have any powers, and her only gadget is her Flip Phone. Instead, she has a natural talent for planning as well as a wide variety of other small talents. She is portrayed by Carly Rose Sonenclar.
- Sigmund Scrambler (played by Mark Linn-Baker) is Annie's uncle, who works as a hypnotist. He is slightly bumbling, but he helps Annie in her pursuit of bothering the Electric Company.
- Sandy Scrambler (played by Ana Gasteyer) is Annie's aunt, who is a hypnotist like Sigmund Scrambler.
- Antigone Carruthers (played by Julie Halston) is Francine's mother, who is the CEO of the Antigone Carruthers Corporation. She usually calls the Electric Company the Electricians. Like Francine, she has an inflated opinion of her own importance, and her idea of punishment for Francine's misdeeds are considered incredibly lenient by the Electric Company.
- Mrs. Bebe Spamboni (played by Andrea Burns) is Manny's mother. She thinks the world of Manny like any mother would, but doesn't approve when he cheats.
- Zandy Rebus is Danny's dog who gave the Electric Company a note in the episode "Pies for puppies."