Paul Reubens as Pee-wee Herman

Joe Manganiello as Joe Manganiello

Jessica Pohly as Pepper

Alia Shawkat as Bella

Stephanie Beatriz as Freckles

Jordan Black as Yul / Party Coordinator (voice)

Doug Cox as Mr. Murgatroyd

Linda Porter as Mrs. Rose

Monica Horan as Ruby the Crossing Guard

Brian Palermo as Marvin

Katherine Vanderlinden as Emily

Tara Buck as Beverly

John H. Mayer as Pharmacist

Richard Riehle as Dan

Dave Power as Dudley

Josh Meyers as Sylvester

Corey Martin Craig as Clovis

Paul Rust as Ernie

Robert R. Shafer as Construction Worker

John Moody as Construction Worker

Daniel Lue as Police Officer

Darrin Henson as Police Officer

Clayton Cannon as Police Officer

Patrick Egan as Gordon the Traveling Salesman

Gloria Vassy as Panicked Female Passenger

Erick Chavarria as Panicked Male Passenger

Frank Collison as Clay

Lynne Marie Stewart as Jimmy

Karen Maruyama as Tourist at Snake Farm

Hal Landon Jr. as Farmer Brown

Michelle Meredith as Linda Brown

Meredith O'Leary as Louise Brown

K.T. Wiegman as Donna Brown

Julie Ouellette as Diane Brown

Alexandra Ella as Janice Brown

Mary Karcz as Betty Brown

Lily Mae Harrington as Pam Brown

Mackenzie Marsh as Judy Brown

Lindsay Hollister as Peggy Brown

Cooper Huckabee as Minister

Darryl Stephens as Rene

Dionne Gipson as Brook

Anthony Alabi as Lane

Sonya Eddy as Wanda

Diane Salinger as Penny King

Brad William Henke as Grizzly Bear Daniels

Leo Fitzpatrick as Abe

Christopher Heyerdahl as Ezekiel

Alan R. Rodriguez as New York City Policeman

Michael Pauley as Guy Who Bumps into Pee-wee

Joseph Mancuso as Pizzeria Employee

David Arquette as DJ

Janina Gavankar as Party Guest

Kelly Bush Novak as Party Guest

Taymour Ghazi as Party Guest

Nicole Sullivan as Shelly

Robert Broski as Abraham Lincoln

Pamela Mant as Queen

Thea Andrews as TV Reporter

John Paragon as TV Cameraman

Charles Robinson as Police Captain

Maya Nalli as Waitress

Deon Aumaier as Man with Toupee (uncredited)

Ryan Nemeth as Construction Worker (uncredited)