The 10K race will start at 10am on Sunday in the city centre with runners heading to Sheffield Wednesday’s Hillsborough stadium before returning.
Organisers said up to 6,000 runners are set to take part and all will be spurred on by live music as part of the Sheffield Bands on the Run festival.
To celebrate Yorkshire’s success at the London Olympics, a custom-made gold medal will be awarded to the 2012th runner across the finish line, while four sterling silver medals are to be awarded to individuals picked at random.
All five medals, which are worth more than £250 each, have been specially produced by Sheffield Assay Office.
Each one will include a special commemorative hallmark celebrating the year of the Queen’s Diamond Jubilee.
A spokesman said: “Yorkshire runners will also be hoping to win the War of the Roses battle against competitors from Lancashire, with every entrant pledging allegiance to their preferred county.
“The winning county will be the one with the fastest average time from their running group.”
Celebrity couple Paula Lane, who plays Kylie Platt in Coronation Street and Tom Shaw, who starred in ITV show DCI Banks are both running for Henshaws Society for Blind People in Harrogate.
The event’s regional charity is Weston Park Hospital Cancer Charity, while the national charity is Diabetes UK.