He played the role of socially awkward IT technician Maurice Moss in Channel 4 sitcom The IT Crowd (2006–2013), for which he won the 2014 BAFTA for Best Male Comedy Performance.

Ayoade has frequently appeared on panel shows, most prominently on The Big Fat Quiz of the Year and served as a team captain on Was It Something I Said?

He has also voiced characters in a number of animated projects, including the films The Boxtrolls (2014), Early Man (2018), The Lego Movie 2: The Second Part (2019), Soul (2020), and The Bad Guys (2022), as well as the series Strange Hill High (2013–2014), Apple & Onion (2018–2021), Krapopolis (2023–present), and Dream Productions (2024).

[8] At the age of 15, he developed an interest in film "beyond Star Wars and Back to the Future" and began exploring the works of directors Woody Allen, Ingmar Bergman, and Federico Fellini.

[11] From 1995 to 1998, Ayoade studied law at St Catharine's College, Cambridge, where he won the Martin Steele Prize for play production[12] and was president of the amateur theatrical club Footlights.

[21] Ayoade was also a writer on the sketch show Bruiser in 2000, which starred former Footlights president David Mitchell and Robert Webb, and featured Holness.

[31][32] In 2007, he directed the music videos for the songs "Fluorescent Adolescent" by Arctic Monkeys and Super Furry Animals's "Run-Away", which starred Matt Berry.

[34] That year he also directed videos for The Last Shadow Puppets songs "Standing Next to Me" and "My Mistakes Were Made for You", the latter of which was inspired by Federico Fellini's Toby Dammit.

[36] Ayoade was featured in Paul King's 2009 film Bunny and the Bull, playing an extremely boring museum tour guide.

The film stars newcomers Craig Roberts and Yasmin Paige with Sally Hawkins, Noah Taylor, and Paddy Considine.

[41][42] Arctic Monkeys and The Last Shadow Puppets frontman Alex Turner contributed five original songs to the soundtrack, inspired by Simon & Garfunkel's music in The Graduate (1967).

The episode pays homage to the 1981 film My Dinner with Andre and was named the "most brilliant half-hour of TV to arrive in this century" by Rolling Stone writer Rob Sheffield.

[45] Ayoade then directed a performance of comedian Tommy Tiernan's world stand-up tour, Crooked Man, which was released in November 2011.

[46] Ayoade provided his voice to the main cast of Channel 4's ill-received animated sitcom Full English, which aired for just five episodes in 2012 before being cancelled.

[48][49] Also in 2012, Ayoade began voicing Todd Lagoona, an anthropomorphic hammerhead shark who was a recurring character in Noel Fielding's Luxury Comedy.

[67] Ayoade made a cameo appearance in the comedy sequel Paddington 2 as a forensic investigator in 2017 and was amongst the voice cast for Vampire Weekend Ezra Koenig's animated series Neo Yokio in the same year.

[75] Ayoade was featured in a supporting role as a pompous artist in both parts of Joanna Hogg's two-part drama The Souvenir.

[76] He lent his voice as a talking ice cream cone to the animated comedy sequel The Lego Movie 2: The Second Part (2019)–which also featured Noel Fielding– and the English version of the Finnish series Moominvalley.

In September 2023, Ayoade received backlash on social media after endorsing the memoir of television writer and anti-transgender activist Graham Linehan, with whom he had worked on The IT Crowd.

"[88] Ayoade is a fan of French New Wave cinema and said in an interview with The Guardian that Louis Malle's Zazie dans le Métro was the film that sparked his interest in filmmaking.

[89] His favourite filmmakers include Malle, Woody Allen, Ingmar Bergman, Orson Welles, Jean-Luc Godard, Jean-Pierre Melville, and Paul Thomas Anderson.

[90][91] Ayoade participated in the 2012 Sight & Sound directors' poll, where he listed his 10 favourite films: The Apartment, Badlands, Barry Lyndon, Crimes and Misdemeanors, Make Way for Tomorrow, Contempt, Ordet, Persona, Raging Bull, and Tokyo Story.

The episode in question was condemned for allowing Fox on as a guest, in particular for when he told a black woman in the audience that discussing racism was "boring".