Zipporah is mentioned in the Book of Exodus as the wife of Moses, and the daughter of Jethro, the priest and prince of Midian.

She is the mother of Moses' two sons: Eliezer and Gershom.

In the Book of Chronicles, two of her grandsons are mentioned: Shebuel, son of Gershom; and Rehabiah, son of Eliezer.

In the Book of Exodus, Zipporah was one of the seven daughters of Jethro, a Kenite shepherd who was a priest of Midian. In Exodus 2:18, Jethro is also referred to as Reuel, and in the Book of Judges (Judges 4:11) as Hobab. Hobab is also the name of Jethro's son in Numbers 10:29.

While the Israelites/Hebrews were captives in Egypt, Moses killed an Egyptian who was striking a Hebrew, for which offense Pharaoh sought to kill Moses. Moses therefore fled from Egypt and arrived in Midian. One day while he sat by a well, Jethro's daughters came to water their father's flocks. Other shepherds arrived and drove the girls away, so that they could water their own flocks first. Moses defended the girls and watered their flocks. Upon their return home, their father asked them, "How is it that you have come home so early today?" The girls answered, "An Egyptian rescued us from the shepherds; he even drew water for us and watered the flock." "Where is he then?", Jethro asked them. "Why did you leave the man? Invite him for supper to break bread." Jethro then gave Moses Zipporah as his wife (Exodus 2:11–21).

After God commanded Moses to return to Egypt to free the Israelites, Moses took his wife and sons and started his journey. On the road, they stayed at an inn, where God came to kill Moses. Zipporah quickly circumcised her son with a sharp stone and touched Moses' feet with the foreskin, saying "Surely you are a husband of blood to me!" God then left Moses alone (Exodus 4:24–26). The details of the passage are unclear and subject to debate.

After Moses succeeded in leading the Israelites out of Egypt, and won a battle against Amalek, Jethro came to the Hebrew camp in the wilderness of Sinai, bringing with him Zipporah and their two sons, Gershom and Eliezer. The Bible does not say when Zipporah and her sons rejoined Jethro, only that after he heard of what God did for the Israelites, he brought Moses' family to him. The most common translation is that Moses sent her away, but another grammatically permissible translation is that she sent things or persons, perhaps the announcement of the victory over Amalek. The word that makes this difficult is shelucheiha, the sendings [away] of her (Ex. 18:2).^{[citation needed]}

Moses' wife is referred to as a "Cushite woman" in Numbers 12. Interpretations differ on whether this Cushite woman [he] was one and the same as Zipporah, or another woman, and whether he was married to them simultaneously, or successively. In the story, Aaron and Miriam criticize Moses' marriage to a Cushite woman. This criticism displeases God, who punishes Miriam with tzaraath (often glossed as leprosy). Cushites were of the ancestry of either Kush (Nubia) in northeast Africa, or Arabians. The sons of Ham, mentioned within the Book of Genesis, have been identified with nations in Africa (Ethiopia, Egypt, Libya), the Levant (Canaan), and Arabia. The Midianites themselves were later depicted at times in non-Biblical sources as dark-skinned and called Kushim, a Hebrew word used for dark-skinned Africans. One interpretation is that the wife is Zipporah, and that she was referred to as a Cushite though she was a Midianite, because of her beauty.