The 30-year-old 'Community' star will bring to life the role he inspired, voicing Miles Morales, a teenager of Black Hispanic descent, who takes over the web-slinging alter-ego following the death of original protagonist Peter Parker, played in the show by Drake Bell, reported USA Today.
The upcoming episode, which will air in the US later this year, will feature Parker travelling to alternate dimensions and meeting other incarnations of Spider-Man.
Glover admits he is flattered to become part of the franchise he adored growing up.
"Spider-Man is the best because you just don't know who he is, and he's funny and he's poor. I understand Spider-Man a lot on that level. He's just trying to make it," he said.