After working as a child actress, Lyonne came to prominence in the late 1990s with her roles in Everyone Says I Love You (1996), Slums of Beverly Hills (1998), But I'm a Cheerleader (1999), and American Pie (1999).

[5] Lyonne was born in New York City,[2] the daughter of Orthodox Jewish parents Ivette Buchinger[2] and Aaron Braunstein, a boxing promoter, race car driver, and radio host.

While in Israel, Lyonne participated in the 1989 Israeli children's film April Fool (Hebrew: אחד באפריל), which began her interest in acting.

[8][22] Lyonne was estranged from her father, who was a Democratic candidate for New York City Council for the sixth District of Manhattan in 2013,[6][23] and lived on the Upper West Side until his death in October 2014.

[25] She was cast as recurring character Opal on Pee-wee's Playhouse at age seven, where she appeared between September and December 1986, and made her film debut that same year with a small part in the Mike Nichols comedy-drama Heartburn.

This led to a headline role in the independent coming-of-age comedy Slums of Beverly Hills (1998), for which she received positive notices for her portrayal of Vivian Abromowitz.

[26] Writing for The Washington Post, Michael O'Sullivan said, "Lyonne is marvelous in conveying Vivian's combination of confusion, curiosity, disgust and desire at what body and psyche are going through.

[30] In the same year, Lyonne played the small but crucial part of Jessica—a role she reprised in two of the film's sequels—in American Pie (1999),[31] which grossed over US$230 million at the box office.

The latter, a follow-up to the 1996 original, was poorly received due to its violence and vulgarity,[33] but Lyonne's portrayal of teenage prostitute Crystal Van Meuther was praised for its "earthy, hard-boiled" nature.

[34] Lyonne played the part of Jeanne, a college activist fighting for lesbian equality, in the acclaimed 2000 television film If These Walls Could Talk 2.

[37] Her portrayal of Debbie Tennis, a psychotic serial killer, in the 2010 horror parody All About Evil was particularly well received, with Film Threat commenting, "[its director] rightfully treats Lyonne as the superstar she is, giving us glimpses of the dark residing in [her] that made Freeway 2: Confessions of a Trick Baby [sic] the final cult masterpiece of the 20th century", noting that "her ability to unleash firehoses of ferocity is on full display here".

Lyonne's work as hard-partying Lou in Antibirth (2016), a psychedelic horror feature inspired by the films of David Cronenberg,[45] drew special attention; Alex McLevy wrote in a review for The A.V.

[46] Other film credits of hers include Sleeping with Other People, Hello, My Name Is Doris, Addicted to Fresno, Hashtag Horror (all 2015); Yoga Hosers, The Intervention (both 2016); Handsome (2017), Show Dogs (2018), Honey Boy (2019), and James Gray's science fiction thriller Ad Astra (2019).

After the final season of Orange Is the New Black, Lyonne began starring as Nadia Vulvokov—a woman trapped in a time loop at her 36th birthday party—on Russian Doll, a comedy-drama series she created and produced along with Leslye Headland and Amy Poehler.

[47] Debuting on Netflix in February 2019, the show was met with rave reviews, with Lucy Mangan of The Guardian calling it "fine [and] impressive," adding, "Nadia is a magnificent creation and Lyonne gives a performance to match".

[80] Regarding her directorial style, Lyonne has expressed frustration with the "simplicity" of modern filmmaking, saying that she likes to counteract this by "filling the frame with an abundance of information", adding: "I do think there's a danger in telling people that brightly lit, crisp things that make perfect sense are good storytelling".

[81] Lyonne has been featured on the covers of magazines including Backstage, Bust, Diva, Glamour, Harper's Bazaar, The Hollywood Reporter, Interview, Nylon, Out, Paper, Variety, Venus, and TheWrap.

[As] much as I love Bette Davis and Mae West and Gena Rowlands, I often found myself identifying with the Peter Falks and the Joe Pescis and the Jimmy Cagneys—all the boys.

[110] In 2005, Lyonne was admitted—under a pseudonym—to Beth Israel Medical Center in Manhattan, suffering from hepatitis C, infective endocarditis, and a collapsed lung; she was also undergoing methadone treatment for heroin addiction.

[19] Her favorite film performances include Giulietta Masina in Nights of Cabiria (1957), Gena Rowlands in A Woman Under the Influence (1974), David Thewlis in Naked (1993), and Roy Scheider in All That Jazz (1979).

[115] During the 2023 WGA strike, she auctioned off the opportunity for fans to solve a New York Times crossword with her to raise money for the Union Solidarity Coalition.