Busco, whose real name was Brandon Moore, died in his sleep on Sunday, his friends and family say.
The dad-of-five gained a following of almost 50,000 on Instagram over the last three years by posting funny clips and short video blogs.
His mum, Valerie Cooper, confirmed the tragic news on Facebook. She posted: "My only child…. My faith will not falter!!!! Lord be my peace."
In a separate post, alongside a picture of her and Busco, devastated Valerie wrote: " I can't believe the earth is still moving when I feel as though my world has stopped…Lord be my peace."
Busco broke onto the scene in 2015 when he jokingly mocked a police officer’s shoes.
As the officer was arresting Busco's pal, the comedian recorded it – and asked "What are those?" in reference to the cop's chunky boots.
After first appearing on Instagram and Vine, the clip went viral, amassing millions of views on social media.
The meme's global popularity spawned thousands of parody versions making fun of people's footwear.
In August 2015, NBA legend Michael Jordan fell victim to the meme at his Flight School summer camp.
During a question-and-answer session, a kid interrupted the basketball star and asked him the catchphrase.
The popular line was even used in Marvel's 2018 film Black Panther.
But Busco was unhappy about the meme's inclusion in the movie.
He told Huffington Post in March that he was "sick" of it and felt "depressed" every time he saw it.
The cause of death is not yet known, but Busco is said to have died just hours after his last joke on social media, People reports.
Tributes have flooded in for the star online, including from his son who wrote that he was the "best dad" he could have wished for.
In his last Instagram post before he died, Busco, true to form, mocked Masion Margiela Fusion Sneakers, which cost $1,645 but are designed to look old and battered.
Reports say a candlelight vigil will be held on Friday at 5.30pm in Berkeley.
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