Maslany was born in Regina, Saskatchewan, Canada, to a translator mother and a woodworker father, the eldest of three siblings. She has two younger brothers. Maslany has Ukrainian, Polish, German, Austrian, and Romanian ancestry.

In 2007, Maslany appeared in The Messengers as Lindsay Rollins. She also starred in the CBC series Heartland where she plays a barrel racer, Kit Bailey, with her horse Daisy. In September 2008, she portrayed Penny, a kidnapping victim in the Canadian television series Flashpoint. Maslany appeared as Sarah Wexlar, the heroin addicted daughter of Dr. Tom Wexlar in the second season of the Canadian comedy drama television series Being Erica in 2010. In 2010, Maslany appeared as the protagonist the Virgin Mary, the mother of Jesus, in the British four part television series The Nativity opposite Andrew Buchan which received excellent ratings.

She appeared in Grown Up Movie Star as Ruby, a role which earned the actor a special jury breakout role award at the 2010 Sundance Film Festival.She played Hannah Simmons in one episode of The Listener. In late 2011, she co-starred with NCIS star Mark Harmon in the film adaptation of John Sandford's Certain Prey as a contract killer named Clara Rinker.

She stars as the lead character, as well as several of her clones, in the 2013 BBC America and Space original series, Orphan Black. The breakout role has earned her legions of fans and the devotion of critics alike.

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