The country music legend revealed that she was recently offered a spot on the ABC reality series, but turned it down without much consideration.
When asked if she'd ever compete on Dancing with the Stars, Parton replied: "Oh no!"
The singer added humorously to The Boot: "They have tried to get me on that ever since it started, even before this movie [Joyful Noise], and I said, 'Are you insane?'"
Jenny McCarthy also recently claimed to have turned down Dancing with the Stars several times.
Dancing with the Stars returns with its new season on March 19 on ABC.
Parton is currently co-starring with Queen Latifah in Joyful Noise, opening on Friday, January 13 in US cinemas.
> Dolly Parton: 'We've been too hard on Miley Cyrus'
> Dolly Parton to work on a musical of her life for Broadway
Watch a trailer for Joyful Noise below: