With fierce rivals Sheffield United waiting in the quarter-final, Stuart Gray's side know victory over their Championship rivals would ensure the first Steel City derby since February 2012.
The original tie was due to take place on February 15 but a waterlogged pitch at Hillsborough saw the match postponed, the second at the ground this season.
Both sides go into the game on the back of morale-boosting league wins on Saturday, Wednesday having beaten Huddersfield 2-0 to end run of two consecutive defeats and Charlton following a 1-0 triumph over promotion-cashing QPR at The Valley.
Johnnie Jackson's late strike on Saturday secured Charlton's third win in 2014 and first in the league, helping their fight against relegation with a crucial three points.
Victory in the cup would see Chris Powell's side book a sixth-round spot for the first time since 2006, although they have been in the last-eight twice since Wednesday's last appearance in 1997.
Gray's men put themselves into the fifth round with victories over Conference side Macclesfield Town and League Two Rochdale, meaning Charlton are the first side from the top two divisions they have faced.
Powell's side knocked League Two Oxford United and Championship rivals Huddersfield Town out of the competition to reach this phase.
The sides have met once in the league this season, with Connor Wickham cancelling out Dale Stephens' opener in a 1-1 draw.
Wickham, who was on loan at Wednesday, has since been recalled by parent club Sunderland, a move that has enabled forward Chris Maguire to step into the spotlight.
The Scot has scored five goals in his last 10 appearances in all competitions, and is the club's top-scorer this season.
