The place and even exact date of his birth are unknown, but he was probably educated in Athens. He was from the Athenian deme of Kudathenaium. He is famous for writing comedies such as The Birds for the two Athenian festivals: the Dionysia and the Lenea. He wrote forty plays, eleven of which still survive, and his plays are the only surviving examples of Old Attic Comedy.

Many of his plays were political, and often satirized the well-known citizens of Athens and their conduct in the Peloponnesian War. He is known to have been prosecuted for Athenian law's equivalent of libel more than once. A famous comedy, The Frogs, was given the unprecedented honor of a second performance. According to a later biographer, he was also awarded a civic crown for "The Frogs".

He appears in Plato's Symposium, giving a humorous mythical account of the origin of Love. The Clouds, a disastrous production resulting in a humiliating and long-remembered (cf. the revised parabasis of "The Clouds" and the parabasis of next year's "The Wasps") last place finish at the City Dionysia, satirizes the new, sophistic learning en vogue among the aristocracy at the time; Socrates was the principal target and in the play he emerges as a typical Sophist, no matter how inaccurate the portrayal may be.

Lysistrata was written during the Peloponnesian War between Athens and Sparta and presents a pacifist theme in a comical manner: the women of the two states show off their bodies and deprive their husbands of sex until they stop fighting. This play was later illustrated at length by Pablo Picasso.