The 49ers played their home games at Jerry Richardson Stadium in Charlotte, North Carolina, and competed in the East Division of Conference USA (C–USA).

All Conference Honorable Mentions:[21] Offense: QB – Chris Reynolds, R-Jr. • RB – Aaron McAllister, R-Sr. • OL – Jaelin Fisher, R-Sr. Defense: DT – Timmy Horne, R-Sr. • DE – Romeo McKnight, R-Sr. • DE – Tyriq Harris, R-Sr. • LB – Tyler Murray, R-Jr. • DB – Ben DeLuca, R-Sr. Special Teams: P – Conner Bowler, R-Jr. 2021 East–West Shrine Bowl:[23] Participants: S - Ben DeLuca • DE - Romeo McKnight 2021 SPIRAL Tropical Bowl:[23] Participant: RB - Tre Harbison 2021 College Gridiron Showcase & Symposium:[23] Participants: RB - Aaron McAllister • OL - Jaelin Fisher Charlotte originally had games scheduled against Norfolk State and Tennessee, which were canceled due to their respective conferences' decisions on the COVID-19 pandemic.

[27] On August 10, fellow Conference USA member Old Dominion suspended all fall sports, creating another opening on the 49ers' schedule.

Several members of the Charlotte offensive line were placed into quarantine following the university's COVID-19 contact tracing protocols, effectively depleting that unit.

[32] On Friday, September 25, Georgia State postponed Charlotte's home opener the next day due to contact tracing issues with a player on their team testing positive for COVID-19.

[45] Marshall canceled the make-up match on December 9, 2020, due to COVID-19 concerns on their team, effectively ending the 49ers season at six games played.

The radio announcers are "Voice of the 49ers" Matt Swierad with play-by-play alongside NFL veteran Al Wallace providing color commentary and Bobby Rosinski and Walker Mehl with sideline reports.

The fog and rain would continue to plague both teams in the nearly empty stadium as Cruz would add another field goal late in the third to bring the score difference to a single digit, but on the next series App stretched the lead back out to eight points only to have Charlotte's Aaron McAllister get it back to within a point on a 97-yard kick off return.

A late touchdown by App made the score difference fifteen points but didn't change the outcome of the game as the 49ers would drop the season opener 20 to 35.

Top performances were dominated by the Mountaineers and included quarterback Zac Thomas who passed for 204 yards and 1 touchdown with 1 interception.

Charlotte would get into the redzone on the final drive of the game but two sacks of Chris Reynolds would lead to time running out as the Niners would lose 17 to 21.

Game Notes: Duke scored quickly and often to defend their home field, going up 24 points before Charlotte could answer with their own first touchdown shortly before the half.

Two devastating fumbles to bookend the second quarter would both result in Western Kentucky touchdowns, putting the 49ers in a sizable scoreless hole going into the half.

Charlotte would find the endzone early in the third quarter to get on the board, but the Hilltoppers would never look back as they held the lead for the entire game.

Due to UNC Charlotte cancelling in-person classes until at least October 1, 2020; it was announced that no attendance would be allowed for the first home game against Georgia State on September 26, 2020.

[33] With the state government announcement at the beginning of October that large venue gatherings could begin at 7% venue capacity, the 49ers announced that for their two October home games (FIU and UTEP) they would allow a maximum capacity in Jerry Richardson Stadium of 1,047 persons, to be limited to players' and coaches' family members, special guests of the university, and students.