A secret Italian government organization takes seriously injured girls, patch them up with artificial body parts, and "conditions" them to be assassins. Each girl is paired with an older man, and they work together as a "fratello" - a brother-sister unit. This moody show takes a look at these girls, and their different relationships with their supervisors. The focus is on Henrietta who is the least conditioned and the one who ponders the most about her newly given life.

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Sequel: Gunslinger Girl: Il Teatrino