The move came after Yiannopoulos led an online troll against Ghostbusters star Leslie Jones who vowed to leave Twitter over her treatment, prompting a major outburst on social media, engadget.com reported on Wednesday.
"People should be able to express diverse opinions and beliefs on Twitter. But no one deserves to be subjected to targeted abuse online, and our rules prohibit inciting or engaging in the targeted abuse or harassment of others," Twitter said in a statement.
"Over the past 48 hours in particular, we've seen an uptick in the number of accounts violating these policies and have taken enforcement actions against these accounts, ranging from warnings that also require the deletion of tweets violating our policies to permanent suspension," it added.
Yiannopoulos reacted to the ban, saying that the site has "confirmed itself as a safe space for Muslim terrorists and 'Black Lives Matter' extremists, but a no-go zone for conservatives."
The Breitbart tech editor has been disturbing Twitter users with his trolls for the past few years. Twitter earlier removed his verified status, but allowed him to keep tweeting.
Last March, Chuck Johnson who once argued that homosexuality caused a horrendous Amtrak train crash, was permanently barred from the site.