When Sarah Ruhl in 2010 first unveiled “Orlando,” her adaptation of the Virginia Woolf novel (subtitled “A Biography”), the subject of gender fluidity had not yet become a cultural obsession. Given Ms. Ruhl’s proclivity for forward-thinking wit and whimsy, though, it seemed logical that the playwright would be drawn to a story — famously inspired by Woolf’s aristocratic lover Vita Sackville-West — about a young man who wakes up one day as a woman.   

For the new revival of “Orlando” that just opened off-Broadway, Ms. Ruhl and director and choreographer Will Davis have enlisted the perfect maverick to play the title character: the celebrated performer and writer Taylor Mac, whose preferred pronoun is judy — though we’ll use they, respectfully, for purposes of this review.

Part of what makes the play charming and affecting is that it explores the assumptions we often make based on gender, and upends them with the quirky, earthy humor and disarming tenderness that are hallmarks of Ms. Ruhl’s writing. Because the play begins in the Elizabethan era, and takes its hero/heroine — whose life spans hundreds of years — into the 20th century and, briefly, the present, this strategy and these qualities are applied to a variety of experiences.

Mx. Mac serves the material with an appealing mix of delicacy and irreverence, at times affirming their perhaps not fully appreciated capacity for understatement. We meet Orlando as a teenager, an aspiring poet who endears himself to the aged queen but then leaves her to begin seducing other members of the opposite sex — women, at this point. 

Orlando becomes engaged, but then is distracted from his fiancée by a Russian princess. He eventually lands at Constantinople, where more debauched encounters pursue; after one, he sleeps for days, then rises to discover he is no longer a gentleman in any sense of that word.

Up until this point — the end of Act One — Mr. Davis keeps things fairly light, even as Orlando ponders death, as fledgling poets must. The six actors who accompany Mx. Mac onstage juggle a variety of roles; Ms. Ruhl provides them with a lot of narration (most of it taken from Woolf, by design), further reinforcing the notion that we are watching a troupe at play.

Janice Amaya’s puckish princess and Nathan Lee Graham’s flamboyant queen are standouts, as is Lisa Kron as a lustful duchess. Oana Botez’s fanciful costume design enhances the air of casual festivity; the shiny flashes of red, green, and gold that first catch the eye suggest a Christmas party.

As “Orlando” follows its protagonist through Victorian England and into the industrial age, the play becomes more poignant, and a bit more provocative. Orlando, who fancied himself a man of action as well as words, laments that as a woman, she “shall never be able to crack a man over the head … or lead an army. … All I can do is to pour out tea and ask my lords how they like it.”  

The female Orlando is courted by an archduke — Ms. Kron again — but feels lonely, until a mysterious man named Marmaduke (a strapping Rad Pereira) arrives on horseback. During their courtship, each of the fledgling partners questions the other’s gender. “You never take more than 10 minutes to dress,” Marmaduke notes of Orlando. “And you listen,” Orlando quips, just as suspicious. 

While Mr. Davis sustains a playful tone overall — transitions between centuries are accompanied by dancing, with the company forming exuberant chorus lines — “Orlando” encourages more and more reflection. The central character’s thoughts turn back to death, but also to poetry, even if we get the impression that Orlando’s talent for the latter remains limited.

The play ends with Orlando professing a renewed vitality and budding clarity. Those may not be traits one associates with the elderly, male or female — let alone someone who has been alive for centuries — but in the fantastical, progressive world that Ms. Ruhl has crafted in Woolf’s honor, anything is possible.