The club was formed in 1873 following a meeting at the towns Spread Eagle Hotel. An announcement in the Gloucester Journal reported "A football club (as rugby was then called) has been formed in this city - the season's operations begin at the Spa on the first Tuesday in next month." A team was then organised to play the College School which was actually played on the ground of the current Kingsholm.

Eleven games were played during the club's 1876-77 season, the club winning 6, losing 2 and drawing 3. Gloucester Rugby Football Club had a successful run in the following two seasons, playing 15 games in both, winning 10, losing 2 and drawing 3 both times. In the 1879-80 season Gloucester RFC played 17 games and lost just two. The 1880-81 season was less successful, winning six of their 13 fixtures.

The club began playing more fixtures as the decade continued, contesting 20 games in the 1884-85 season, and up to 34 in the 1891-92 season. In 1891, after argument with the cricket club, Gloucester left the Spa (the ground they were sharing) because, during the winter the Rugby Club had used a salt mixture to remove frost from the pitch, resulting in the death of the grass on the wicket. Gloucester RFC were no longer welcome at the Spa ground. They then acquired lands from the Castle Grim Estate for £4,000 and have resided in this place, known since as Kingsholm.

Gloucester RFC were suspended by the RFU for professionalism as a result of the club recruiting players: the "Shewell case". The club responded by adopting a more puritanical adherence to the amateur regulations, and were reinstated. However when the Northern clubs split from the RFU to form the Northern Rugby Union (later known as the Rugby Football League) a number of Gloucester RFC players "went North".

The 1920-21 season became famous for Gloucester RFC when Fred Webb skippered the club to 23 matches unbeaten at home at Kingsholm, with only the United Services and Pontypool clubs being able to defeat them at home. The following season was infamous for the number of footballers that were sent from the field by officials, 14 for fighting, seven for arguing, six for language and one for foul play. Leicester actually cancelled their Gloucester RFC fixture because of this. Around the mid 1920s the media began to refer to the club as the 'Red and Whites'.

In 1972 Gloucester RFC won the first ever National Knock-Out Competition. Having overcome Bath, Bristol, London Welsh and Coventry (all away from home) in earlier rounds, they beat Moseley in a Twickenham final that was marred by violence and the sending off of Moseley's Nigel Horton.

In 1978 Gloucester RFC won the first ever John Player Cup, defeating Leicester in another final noted for violent play both on and off the pitch at Twickenham.

Despite the two cup wins of the 1970s and a shared trophy in 1982, the Tigers found themselves in the shadow of Bath. Bath finished runners-up in the league in 1989, but the disastrous end to the 1989-90 campaign was to be a prelude to lean years. Closing in on English rugby's first 'Double', Gloucester RFC's last-day collapse handed Wasps the league title before a 48-6 cup final heartbreak at the hands of Bath.

However, te 1992-93 season was a different story. The players called a crisis meeting with club coach Keith Richardson about the apparent lack of player rewards relative to other comparable clubs. A year later, club chairman Peter Ford restated the club's traditional anti-professionalism: "We play by the rules. .. whatever anybody else does, we'll stick by the rules. If they say we can't pay players, or offer them inducements or cars or flats. .. then we won't do it."

When professionalism came in 1995, Gloucester RFC was without a major investor and lost ground in terms of player recruitment and revenue acquisition. However, this did not prevent the club from transforming itself into a limited company.

In April 1997, Tom Walkinshaw became the club’s new owner with a majority shareholding. Richard Hill's position would soon be filled by former France captain Philippe Saint-André.

In Saint-Andre's first full season, a third-place finish in 1999-2000 took Gloucester into the Heineken Cup. With Phil Vickery, Trevor Woodman and All Blacks legend Ian Jones forming the basis of a formidable pack, Gloucester RFC marched their way to the last four, where only last-ditch Leicester defending would deny them a final appearance.

Saint-Andre's acrimonious departure couldn't stop them winning their first cup triumph in 25 years in 2003, under new coach Nigel Melville.
Unstoppable in the league, Gloucester RFC reached the inaugural Championship final 15 points clear of their nearest rival, however under the new league system Gloucester RFC were 'rewarded' with a 3 week lay-off, while Wasps played regular rugby during the 3 weeks as a result Gloucester RFC lost their momentum and were crushed by Wasps at Twickenham.

In the 2004-05 season Gloucester failed to qualify for the Heineken Cup. Consequently, Nigel Melville left the club and was replaced by Dean Ryan for the 2005-06 season.

At the start of the 2005 season, the club chairman Tom Walkinshaw began several changes to modernise the club in order to keep up with rugby's rapidly advancing professionalism, whilst the majority of fans supported the need to modernise, some fans were concerned that the club was losing touch with its heritage and traditions. 'Gloucester Rugby Football Club' was renamed 'Gloucester Rugby' and, due to copyright issues, it no longer used Gloucester City's Coat of Arms as the clubs crest (as the crest didn't belong to the club, and with no copyright, unofficial merchandise was freely available).

The 2005-06 season was an improvement in the clubs fortunes, although they did not qualify for the play-offs, they were strong contenders and lost out on the last day of the regular season. They also won silverware in the European Challenge Cup, defeating London Irish in a tense final that went into extra time. This season was also a glimpse of the incredible talent Gloucester RFC were bringing up from their academy.

In 2006-2007 Gloucester Rugby finished 1st in the Guinness Premiership table, both Leicester and Gloucester Rugby tied with 71 points, Gloucester Rugby gaining first place with more games won, they also demonstrated their level of skill and vision in the Heineken Cup, against sides such as Leinster and Edinburgh despite not progressing past the group stage, Gloucester Rugby defeated the Saracens in the semi-final at Kingsholm, 50-9, to move into the Twickenham final where they faced Leicester Tigers. However, Gloucester Rugby were beaten heavily by the Tigers with the final score being 44-16, Gloucester Rugby again being defeated by the play-off system.

Gloucester Rugby began the 2007-08 Guinness premiership campaign as favourites, and started the season strongly winning their first five games before defeat away to London Irish. Dean Ryan's main focus for the 2007-08 season was for Gloucester to establish themselves in Europe, but despite coming top of their group they lost to Munster in the Quarter Final. After the Heineken Cup exit Gloucester overcame their poor mid-season form with a series of victories capping it off with their first away win against Wasps for 18 years, and then defeating Bath in an intense encounter at Kingsholm to win the league for the second year running, and book a home semi-final in the Championship Play-off against Leicester Tigers in a repeat of last seasons Championship final. The result was the same, although a closer affair, the Tigers won 25-26.

After failing to qualify for the play-off's in 2008/09 and defeat to Cardiff in the EDF Energy Cup Final at Twickenham, Gloucester announced on 11 June 2009 that Dean Ryan had left the club by mutual consent. He was replaced by assistant Bryan Redpath.