The teams have been drawn in Group G of the qualifiers, where they will do battle with Mali and Congo for a place in next year's showpiece.
Match preview
Gambia played out a hard-fought 1-1 stalemate against the United Arab Emirates in Sunday's meeting at the Zabeel Stadium in Dubai.
Tom Saintfiet's men trailed to Ali Mabkhout's penalty seven minutes shy of half time before restoring parity in the early exchanges of the second period thanks to Musa Barrow.
The Scorpions now prepare for their journey towards another AFCON appearance having seen off the threat of Chad in the preliminary qualifying phase.
Lying in store next is the challenge of an opposing side who suffered a 2-1 defeat in the teams' only previous encounter last year.
Having lost just one of eight games since the turn of the year, Saturday's hosts will back themselves to get the job done and take a step in the right direction towards securing progress into the group stages of the upcoming continental finals.
South Sudan made light work of Djibouti in the previous round of the qualifiers, where they claimed a 5-2 victory on aggregate.
A 4-2 first-leg victory on March 27 set Stefano Cusin's men on their way before a 1-0 triumph in the return leg four days later completed the job.
The Bright Stars, who lost all of their five matches in 2021, have now begun 2022 with back to back victories, which bodes well for what is to come.
Still awaiting their maiden appearance at the AFCON, Saturday's visitors set out to pick up where they left off during the last international break.
Gambia form (all competitions):
South Sudan form (all competitions):
Team News
Gambia's Assan Ceesay was at the double in his last game against this weekend's opponents, making him one to keep an eye on.
Hamza Barry has returned to the international scene for the first time since 2019 when he suffered a long-term knee injury.
Meanwhile, Pa Modou Jagne has been left out of his side's latest squad and will not have the opportunity to extend his lead at the top of the all-time appearances chart.
Nineteen-year-old William Gama will be aiming to deliver the goods once again having recently opened his national-team account at the sixth time of asking.
Michael Marco still awaits his first cap for the away side, but Juma Genaro is expected to retain his place in goal for a record-extending 31st international outing.
Gambia possible starting lineup:
Jobe; Colley, Sanneh, Touray, Gomez; Marreh, Jallow, Bobb; Ceesay, Barrow, Jobe
South Sudan possible starting lineup:
Genaro; Maker, Abdelkarim, Sebit, Toha; Chol, Morgan, Musa; Okello, Emmanuel, Majak