John Wilkes Booth was born on May 10, 1838 on a small farm known as Tudor Hall, located a few miles outside of Bel Air, Harford County, Maryland. His parents, Junius Booth and Mary Ann Holmes were British, and moved to the United States in 1821. John Wilkes was the ninth born of their ten children. The Booth family stayed on their farm at Tudor Hall in the warmer months, and also owned and used slaves. During the winter, they would move to their other home located on North Exeter Street in Baltimore City. His father, Junius was a very well-known actor.

John Wilkes received his schooling in Maryland, first at a boarding school in Cockeysville (Baltimore County) operated by Quakers, after which he attended St. Timothy's Hall, an Episcopal military academy in Catonsville, also in Baltimore County. As a young man in the 1850s, Booth joined the Know-Nothing Party. This party was formed by Americans for the purpose of preserving the country for native-born white citizens. In 1852, his father died and John left school and spent the next few years working on their farm near Bel Air.

However, farming was not in John's life plan. According to his sister, Asia Booth Clarke, he had always wanted to be famous. So, John followed in his father's footsteps toward a career in acting. In 1855, at the age of seventeen, Booth made his acting debut in the production of Shakespeare's Richard III, in the role of the Earl of Richmond. Following this, it would be two years before he again appeared on stage. He did play minor roles in Philadelphia, but was not well received, as he would frequently forget his cues and lines. But, desiring to be a good actor, he did not give up, and finally landed roles in 1858 as part of the Richmond Theatre. Living in Richmond, he became fascinated with the people and the Southern way of life. In 1859, Booth witnessed the execution of John Brown, working as one of the guards stationed there to prevent any attempts to rescue Brown.

In 1860, his acting career started to take off. He landed the role as Duke Pescara in The Apostate, at the Gayety Theatre in Albany, New York. It was here that President Lincoln passed through Albany en route to Washington, D.C. Booth's acting continued in such productions as Romeo and Juliet, The Marble Heart, The Merchant of Venice, Julius Caesar, Othello, The Taming of the Shrew, Hamlet, Macbeth, and others. His appearances took him to New York, Boston, Baltimore, Washington, Cleveland, Chicago, Detroit, St. Louis, Leavenworth, Nashville, New Orleans and Richmond, prior to the Civil War. From November 2 through November 15, 1863, Booth appeared in The Marble Heart in the role of Raphael at Ford's Theatre in Washington, D.C. It was during his November 9 performance that President Lincoln attended and saw Booth in this role. The box where Lincoln sat was the same exact spot in which he would be later assassinated. Following The Marble Heart at Ford's, Booth made only one more appearance there, when on March 18, 1865, he appeared as Duke Pescara in The Apostate. Although Booth was considered a good actor, he never excelled to the level of talent possessed by his father, nor his brother Edwin, who all worked together in one production of Julius Caesar. In the play, Booth appeared as Marc Antony, while Edwin played Brutus, and their father Julius, played Cassius. In the summer of 1864, Booth appeared in a production at Meadville, Pennsylvania, and stayed in a room at the McHenry House. Upon checking out, a cleaning woman attending the room found an inscription on one of the windowpanes that read, "Abe Lincoln departed his life August 13, 1864, by the effects of poison." Unfortunately, no one gave it much attention, nor focused on Booth as the writer.

Late in the summer of 1864, Booth began his plans to kidnap Abraham Lincoln. His idea was to abduct Lincoln, then take him to Richmond where he would be held for ransom in exchange for Confederate prisoners held in Union camps. Booth believed this was one way to increase the deteriorating ranks of Rebel soldiers. To assist him in his plan, he began recruiting helpers. He was able to recruit Michael O'Laughlen, Samuel Arnold, Lewis Paine, John Surratt, David Herold and George Atzerodt. A meeting was set whereby all the members met at Gautier's Restaurant, a short distance from Ford's Theatre in Washington, and discussed their abduction plan.

A few days after this meeting, Booth became aware that Lincoln would be attending a play at the Campbell Hospital near Washington, D.C., on March 17, 1865. To Booth this seemed to be his best opportunity to abduct Lincoln. That day in March came and Booth's plan was in place. Lincoln decided at the last minute not to attend the play, but rather to speak to the 140^th Indiana Regiment and present a captured Confederate flag to the Governor of Indiana. As this plan failed, some of Booth's conspirators became disenchanted and abandoned the group.

In Booth's personal life was a lady named Lucy Lambert Hale, the daughter of John Parker Hale, an abolitionist senator from New Hampshire. In early 1865, the Hale family moved into the National Hotel, in Washington, where Booth was also staying. In the first days of March 1865, Booth became secretly engaged to Lucy. On March 4, on the occasion of Lincoln's second inauguration, Booth invited Lucy as a guest. In his quest for Lincoln, Booth once confided to an actor friend by the name of Samuel Knapp Chester, that, "What an excellent chance I had to kill the President, if I had wished, on inauguration day!" Booth and his conspirators had other plans to kidnap Lincoln, specifically at a theatre, but these plans fell through. Then on April 9, 1865, General Robert E. Lee surrendered the Army of Northern Virginia to General Grant, effectively ending the Civil War. Two days later, President Lincoln gave a speech at the White House where he discussed possible new rights for blacks. In the audience were Booth, Herold and Paine. The speech enraged Booth, and he vowed to silence Lincoln.