She attended Interlochen Arts Academy, the Yale School of Drama and the North Carolina School of the Arts. Margot began her career in 1980 working in theater and television commercials and was a member of the original company of I'm Getting My Act Together & Taking it on the Road at the New York Shakespeare Festival, then later at the Circle in the Square Downtown.

Of her performance in the 1996 theatrical production Sordid Lives Variety wrote "Also lending solid support are ... and Margot Rose as the well-worn Betsy Mae, whose gentle musical offerings of such ballads as 'Will the Circle Be Unbroken,' 'The Water Is Wide' and an original title song punctuate the onstage doings."

Of Rose's performance in the theatrical production of The Gay '90s Musical in 1997, Variety wrote the play "features the beautifully harmonized voices of Margot Rose, [Bill] Ledesma and [Bill] Hutton as a trio of gay friends who nurture and support each other throughout their lives".

She has guest starred in over sixty television series including: Hill Street Blues, E/R, Night Court, Star Trek: The Next Generation, Star Trek: Deep Space Nine, L.A. Law, The West Wing, Judging Amy, Desperate Housewives, Law and Order: Los Angeles, Murphy's Law, The Mentalist and Numb3rs, among many others. She has also had a number of roles in films that include: 48 Hrs., A Civil Action, True Believer, Brewster's Millions and Hollow Man. Rose composed the music for the 2000 film Sordid Lives and the 2008 television series Sordid Lives: The Series. She also performed in the original Del Shores stage production of Sordid Lives in 1996 in the role of Bitsy Mae Harling.