Batman, Batgirl, and Robin forge an alliance with the Teenage Mutant Ninja Turtles to fight against the Turtles' sworn enemy, The Shredder, who has apparently teamed up with Ra's Al Ghul and The League of Assassins.
| Actor | Role |
|---|---|
 Troy BakerBatman / Joker (voice) | Batman / Joker (voice) |
 Eric BauzaLeonardo (voice) | Leonardo (voice) |
 Darren CrissRaphael (voice) | Raphael (voice) |
 Kyle MooneyMichelangelo (voice) | Michelangelo (voice) |
 Baron VaughnDonatello (voice) | Donatello (voice) |
 Carlos AlazraquiBane (voice) | Bane (voice) |
 Cas AnvarRa's al Ghul (voice) | Ra's al Ghul (voice) |
 Rachel BloomBatgirl (voice) | Batgirl (voice) |
 John DiMaggioMr. Freeze (voice) | Mr. Freeze (voice) |
 Keith FergusonBaxter Stockman / Two-Face (voice) | Baxter Stockman / Two-Face (voice) |
 Brian GeorgeAlfred (voice) | Alfred (voice) |
 Ben GirouxRobin (voice) | Robin (voice) |
 Tom KennyPenguin (voice) | Penguin (voice) |
 Andrew KishinoShredder (voice) | Shredder (voice) |
 Jim MeskimenCommissioner Gordon / Scarecrow (voice) | Commissioner Gordon / Scarecrow (voice) |
 Tara StrongHarley Quinn / Poison Ivy (voice) | Harley Quinn / Poison Ivy (voice) |