He began his career in music in January of 1977 as a roadie for the punk rock band The Ravers (who would later change their name to The Nails). In the fall of that year, he began attending the University of California, Santa Cruz. He spent one quarter of the year studying acting and the history of Paraguay before leaving to become involved in San Francisco, California's punk scene. In June of 1978 he responded to an ad put out by guitarist East Bay Ray and together they formed the Dead Kennedys. He began performing with the band under the stage name Occupant, but shortly after began using his current stage name. Biafra wrote the band's lyrics, most of which were political in nature and displayed a sardonic, sometimes absurdist, sense of humor despite their serious subject matter. In June of 1979, Biafra co-founded the record label Alternative Tentacles with which the Dead Kennedys released their first single, "California Über Alles". The label was created to allow the band to release albums without having to deal with pressure from major labels to change their music (although the major labels were not willing to sign the band due to their songs being deemed too controversial). After dealing with Cherry Red in the UK and IRS Records in the US for their first album Fresh Fruit for Rotting Vegetables, the band released all later albums (and later pressings of Fresh Fruit) on Alternative Tentacles (with the exception of live albums released after the band's break-up, which the other band members compiled from recordings in the band partnership's vaults without Biafra's input or endorsement). Biafra has been the owner of the company ever since its founding, though he does not receive a salary for his position (Biafra refers to his position in the company as the "absentee thoughtlord").

In the fall of 1979, Biafra ran for mayor of San Francisco as a prank, using the Jello ad campaign catchphrase, "There's always room for Jello", as his campaign slogan. Having entered the race before creating a campaign platform, Biafra later wrote his platform on a napkin while attending a Pere Ubu concert. As he campaigned, Biafra wore campaign t-shirts from his opponent Quentin Kopp's previous campaign and at one point vacuumed leaves off the front lawn of another opponent, current U.S. Senator Dianne Feinstein, to mock her publicity stunt of sweeping streets in downtown San Francisco for a few hours. Supporters committed equally odd actions; two well known signs held by supporters said "If he doesn't win I'll kill myself" and "What if he does win?" His platform included unconventional points such as forcing businessmen to wear clown suits within city limits and a citywide ban on cars (although the latter point was not considered abnormal by many voters at the time, as the city was suffering from serious pollution problems). Biafra has expressed irritation that these parts of his platform attained such notoriety, preferring instead to be remembered for serious proposals such as legalizing squatting in vacant, tax-delinquent buildings and requiring police officers to keep their jobs by running for election voted on by the people of the neighborhoods they patrol. During a TV interview (reproduced on The Early Years home video and DVD), he had this to say to those who thought less of him running for mayor: “For those of them who have seen my candidacy as a publicity stunt or a joke, they should keep in mind that it is no more of a joke, and no less of a joke than anyone else they care to name.” He finished fourth out of a field of ten, receiving 3.5% of the vote (6,591 votes); the election ended in a runoff that did not involve him (Feinstein was declared the winner). In reaction to his campaign (and that of Sister Boom-Boom, a drag queen who also ran for mayor and handily won the third place spot above Biafra), San Francisco passed a resolution stating that no candidate could run under any name other than their given name.

The Dead Kennedys toured widely during their career, starting in the late 1970s. They began playing mostly at southern Californian clubs (most notably the Whisky a Go Go), but eventually they moved on to major clubs across the country, including the CBGB in New York. Later, they played to larger audiences such as at the 1980 Bay Area Music Awards (where they played the notorious "Pull My Strings" for the first and only time), and headlined the 1983 Rock Against Reagan festival.

Biafra married Therese Soder, aka Ninotchka, lead singer of San Francisco-area punk band The Situations on October 31, 1981. Flipper vocalist/bassist Bruce Loose conducted the wedding, having paid to join the Universal Life Church as a minister just to conduct the ceremony, which took place in a graveyard. The wedding reception, which members of Flipper, Black Flag, and D.O.A. attended, was held at director Joe Reis' Target Video studios. The marriage ended in 1986.

Biafra became a spoken word artist in January 1986, starting with a performance at University of California, Los Angeles. In his performance he combined his sense of humor with his political beliefs, much in the same way that he did with the lyrics to his songs. Biafra has held this career since, but did not begin recording spoken word records until after the disbanding of the Dead Kennedys.

In 1999, Biafra and other members of the anti-globalization movement protested the WTO Meeting of 1999 in Seattle. Along with other famous musicians from the west coast, he formed the short-lived band the No WTO Combo to help promote the movement's cause. The band was originally scheduled to play during the protest, but the performance was canceled due to riots. The band performed the following night at the Showbox in downtown Seattle (outside of the curfew area) where they played a short set. The hiphop group Spearhead also played at the event. A CD containing recordings from the concert, titled Live from the Battle in Seattle, was later released.

In 2000, the New York State Green Party drafted Biafra as a candidate for the Green Party presidential nomination, and a few supporters were elected to the party's nominating convention in Denver, Colorado. Despite the fact that his address to the convention was positively received, the party overwhelmingly chose Ralph Nader as the presidential candidate. Nader and the Green Party have grown increasingly popular among the punk rock community with artists like Patti Smith, Propagandhi, Thought Riot, Hungry March Band, and Partyline, among others.

Biafra, along with a camera crew (dubbed by Biafra as "The Camcorder Truth Jihad"), later reported for the Independent Media Center at the Republican and Democratic conventions. Biafra detailed these events in his album Become The Media, which has resulted in him being credited with coining the slogan "Don't hate the media, become the media". Indymedia and related alternative media often use this line, or the now more apt "Don't hate the media, be the media."

Also in 2000, Biafra gave the keynote speech at the H.O.P.E. 2000 ("H2K") hacker conference. Though Biafra had never used a computer in his life, the attendants saw him as being capable of drawing insightful connections between hacking and activism. He has continued to deliver keynote speeches at every H.O.P.E. conference since, the most recent of which began with him mooning the audience. [3] Audio of these speeches are freely available online for download.

In April of 2001, Biafra took part in a protest against the Free Trade Agreement of the Americas conference in Quebec.

His eighth spoken word album, In the Grip of Official Treason, was released in October of 2006.

In 1988, Biafra and Alain Jourgensen of the band Ministry formed the band Lard. The band became a side project for the members of Ministry, with Biafra providing vocals and lyrics. While working on the film Terminal City Ricochet in 1989, Biafra did a song for the film's soundtrack with D.O.A.. As a result, Biafra worked together with D.O.A. on the album Last Scream of the Missing Neighbors. Biafra also worked with Nomeansno on the soundtrack, which lead to their collaboration on the album The Sky is Falling and I Want My Mommy the following year.

In 2005 Biafra appeared on Episode 285 of This American Life, themed "Know Your Enemy", which featured a phone call between Jello Biafra and Michael Guarino, the prosecutor in the Frankenchrist trial. The episode was about Guarino's change of opinion and the reconciliation between Guarino and Biafra.

On May 7, 1994 people who believed Biafra was a sell out attacked him at the 924 Gilman Street club in Berkeley, California. Biafra claims that he was attacked by a man nicknamed Cretin, who crashed into him while slamdancing. The crash injured Biafra's leg, causing an argument between the two men. During the argument, Cretin pushed Biafra to the floor and five or six friends of Cretin assaulted Biafra while he was down, yelling "Sellout rock star, kick him". Biafra was later hospitalized with serious injuries, the attack derailed Biafra's plans for both a Canadian spoken-word tour and an accompanying album

As of late 2005, Biafra currently performs with the band The Melvins. The new band was dubbed "Jello Biafra and the Melvins", though fans often refer to them as "The Jelvins." Together they have released two albums.