Stine was born on October 8, 1943 in Columbus, Ohio, the son of Lewis Stine, a shipping clerk, and Anne Feinstein. He grew up in Bexley, Ohio. His family was Jewish. He began writing at age nine, when he found a typewriter in his attic, subsequently beginning to type stories and joke books. He graduated from Ohio State University in 1965 with a Bachelor of Arts in English. While at OSU, R. L. Stine edited the OSU humor magazine The Sundial for three out of his four years there. He later moved to New York City to pursue his career as a Writer.

On June 22, 1969, Stine married Jane Waldhorn, an Editor and Writer who later co-founded Parachute Press in 1983. The couple's only child, Matthew (born June 7, 1980) works in the music industry.

Stine wrote dozens of humor books for kids under the name Jovial Bob Stine and created the humor magazine Bananas. Bananas was for teenagers published by Scholastic Press for 72 issues between 1975 and 1984, plus various "Yearbooks" and paperback books. Stine was Editor and responsible for much of the writing (other contributors included Writers Robert Leighton, Suzanne Lord and Jane Samuels and artists Sam Viviano, Samuel B. Whitehead, Bob K. Taylor, Bryan Hendrix, Bill Basso, and Howard Cruse). Recurring features included "Hey – Lighten Up!", "It Never Fails!", "Phone Calls", "Joe" (a comic strip by John Holmstrom), "Phil Fly", "Don't You Wish...", "Doctor Duck", "The Teens of Ferret High", "First Date" (a comic strip by Alyse Newman), and "Ask Doctor Si N. Tific".

In 1986, Stine wrote his first horror novel, called Blind Date. He followed with many other novels, including The Babysitter, Beach House, Hit and Run, and The Girlfriend. He was also the co-creator and head Writer for the Nickelodeon Network children's television show Eureeka's Castle, original episodes of which aired as part of the Nick Jr. programming block during the 1989-1995 seasons.

In 1989, Stine started writing Fear Street books. Before launching the Goosebumps series, Stine authored three humorous science fiction books in the Space Cadets series titled Jerks in Training, Bozos on Patrol, and Losers in Space. In 1992, Stine and Parachute Press went on to launch Goosebumps.

Also produced was a Goosebumps TV series that ran for four seasons from 1995–1998 and three video games; Escape from HorrorLand, Attack of the Mutant and Goosebumps HorrorLand. In 1995, Stine's first novel targeted at adults, called Superstitious, was published. He has since published three other adult-oriented novels: The Sitter, Eye Candy, and Red Rain.

According to Forbes List of the 40 best-paid Entertainers of 1996–97, Stine placed 36th with an income of $41 million for the fiscal year. His books have sold over 400 million copies worldwide as of 2008, landing on many bestseller lists. In three consecutive years during the 1990s, USA Today named Stine as America's number one best-selling author.

Among the awards he has received are the 2002 Champion of Reading Award from the Free Public Library of Philadelphia (that award's first year), the Disney Adventures Kids' Choice Award for Best Book-Mystery/Horror (three-time recipient) and the Nickelodeon Kids' Choice Awards (also received three times). During the 1990s, Stine was listed on People Weekly's "Most Intriguing People" list, and in 2003, the Guinness Book of World Records named Stine as the best-selling children's book series author of all time. He won the Thriller Writers of America Silver Bullet Award in 2007, and the Horror Writers Association's Lifetime Achievement Award in 2014. His stories have even inspired R. L. Stine's Haunted Lighthouse, 4D movie-based attractions at SeaWorld (San Antonio and San Diego) and Busch Gardens (Williamsburg and Tampa).

In the first decade of the 21st century, Stine has worked on installments of five different book series, Mostly Ghostly, Rotten School, Fear Street, The Nightmare Room, Goosebumps Horrorland and the stand-alone novels Dangerous Girls (2003) and The Taste of Night (2004). Also, a direct-to-DVD movie The Haunting Hour Volume One: Don't Think About It, starring Emily Osment was released by Universal Home Entertainment on September 4, 2007. Jack Black portrayed a fictionalized version of Stine in the 2015 film Goosebumps, while Stine himself made a cameo appearance, playing a Teacher named "Mr. Black".

Stine has been referred to as the "Stephen King of children's literature" and is the author of hundreds of horror fiction novels, including the books in the Fear Street, Goosebumps, Rotten School, Mostly Ghostly, and The Nightmare Room series. Some of his other works include a Space Cadets trilogy, two Hark gamebooks, and dozens of joke books. As of 2008, Stine's books have sold over 400 million copies.

Both were re-released in 2010 under the name Bitten. ISBN 978-0060530808