Zoe Ann Lyons was born on 3 October 1971 in Haverfordwest, Pembrokeshire, Wales, to an Irish father and an English mother. Her family soon moved to Ireland, where she attended a Catholic primary school in Dunmore East, County Waterford. The family then moved to Clonmel, County Tipperary, then to Epsom, Surrey when Lyons was nine, and then to Glasgow. Her first job was in a jam factory in Glasgow.

Zoe initially studied psychology at the University of York, graduating in 1992. She soon developed a love for performing, moving to London to train at The Poor School and Workhouse Theatre before taking to the stage in dramatic and comedic roles. Her first foray into comedy took place around 2003.

In 2021, Lyons hosted her own teatime TV quiz show Lightning on BBC Two, as well as Stand Up and Deliver on Channel 4, where she mentored actress Katie McGlynn in stand-up comedy. Lightning combined elements of the traditional game show and quiz show, with six contestants battling across frenetic rounds answering trivia questions and undertaking physical trials, playing for an ultimate prize of £3,000.

Lyons lives in Brighton with her partner Sindy, a nurse originally from the Netherlands. Lyons shared an Instagram post in 2020 about her alopecia in the hope that it would help others. The couple sold their flat after dealing with structural issues and relocated to nearby Hove, where they live with their Jack Russell, Groucho Barx.