Winona LaDuke was born in Los Angeles, California to Vincent and Betty LaDuke. Her father was part Anishinaabe (Ojibwe or "Chippewa") from an Indian reservation of Minnesota. He was an actor with supporting roles in Western movies, an activist, a writer, and at the end of his life, a spiritual guru under the name Sun Bear. Her mother was a Jewish artist, employed as an art professor at Southern Oregon University in Ashland, Oregon. LaDuke is the mother of five.

LaDuke was raised in Ashland, Oregon, but after graduating from Harvard in 1982 with a degree in rural economic development, she accepted a job as principal of the high school on the Anishinaabe White Earth Indian Reservation in Minnesota. She then graduated with an M.A. in Community Economic Development from Antioch University. She soon became an activist, involved in the struggle to recover lands promised to the Anishinaabe by an 1867 treaty. She helped the Anishinaabe buy back thousands of acres of ancestral land.[citation needed]

She worked with Women of All Red Nations to publicize the alarmingly high level of forced sterilization among Native American women.

LaDuke was named Woman of the Year by Ms. Magazine in 1997 and won the Reebok Human Rights Award in 1998. She is the founder of the White Earth Land Recovery Project in Minnesota and the Indigenous Women's Network. She is also Executive Director of Honor the Earth, an organization she co-founded with Indigo Girls in 1993. The Native-led organization's mission is "to create awareness and support for Native environmental issues and to develop needed financial and political resources for the survival of sustainable Native communities. Honor the Earth develops these resources by using music, the arts, the media, and Indigenous wisdom to ask people to recognize our joint dependency on the Earth and be a voice for those not heard."

LaDuke is the author of the novel Last Standing Woman (1997), the non-fiction book All our Relations: Native Struggles for Land and Life (1999), and Recovering the Sacred: the Power of Naming and Claiming (2005), a book about traditional beliefs and practices.