In the spiritual, the group of people that are oppressed are the ancient Israelites. The opening stanza of the song depicts the Jewish slaves who were oppressed by the Pharaoh of Egypt. This is a retelling of the book of Exodus in which Moses is chosen by God to deliver the Hebrew slaves from bondage at the hands of the Egyptians. "Let my people go" is the cry that Moses gives to Pharaoh in the demand that Pharaoh alleviate the suffering of the Jewish people who were oppressed. The song's focus is the deliverance of this oppressed group from bondage. In the spiritual, the use of the Exodus narrative becomes essential in understanding the condition of enslavement.
For the slave in America, the song acquires significance when seen in this Biblical light. The cries of the oppressed in the song matches the condition of the slave in America. In this context, evil Pharaoh would be the slaveowner in the South, while the cry of "let my people go" would refer to the African enslaved in America. The song works in the context of the slavery because so many slaves envisioned the situation between the modern slave and slaveowner as reflective of the Biblical condition of the ancient Israelites and the Egyptian Pharaoh. The resonating cry of "let my people go" and the invocation of Moses, who freed the Israelites from the clutches of enslavement, help to enhance the meaning of the song in the application to the predicament of slavery in the American modern setting.
Another connection between the oppressed people in the song and the modern slave is the call to Moses. In the song, Moses comes and delivers those who are enslaved from bondage. The context of slavery saw Harriet Tubman, known as "Black Moses," as this force of deliverance. The call to Moses in the song parallels the call out to Harriet Tubman, who used the Underground Railroad to free hundreds of slaves. In this, the spiritual spoke of redemption and salvation in the future, but also spoke of how this could be in the present in the form of escape into freedom.