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Midnight Sparkle a.k.a Human Twilight Sparkle is the main antagonist of Mario meets Twilight Sparkle. She is voiced by Tara Strong who also voiced Twilight Sparkle in her pony form.

Midnight Sparkle is mentioned by Twilight Sparkle when she and Spike told Mario and Luigi about Canterlot High and says that her evil side is going to destroy the high school by getting the magic from Equestria a.k.a her town. Twilight, Mario, Luigi, Emmet, and Spike decide to go to Canterlot High and stop Twilight's human form from unleashing the magic and turning into Midnight Sparkle before is too late.

Midnight Sparkle 2