O'Keefe, 36, on Tuesday, stated he was publishing audio recordings of several network meetings which he insists will 'expose' Zucker and other CNN executives declaring their 'political biases'.
The calls O'Keefe had been recording for two months now are the network's daily 9 am editorial meeting with senior staff from CNN's main bureaus, which is run by Zucker.
The move by Project Veritas is the latest sting operation organized upon the news station by the conservative operative who has long-targeted CNN over allegations of liberal and anti-Trump bias.
O'Keefe taunted the exposé on Twitter by sharing a live stream recording of the moment he ambushed Zucker throughout a phone call telling him he was preparing to make the footage of his rundowns public.
'Mr Zucker, are you there? Hey, this is James O'Keefe. We've been listening to your CNN calls for basically two months, and recording everything. I just wanted to ask you some questions if you have a minute,' he started.
'Do you still feel you're the most trusted name in news, because I have to say from what I've been hearing on these phone calls, I don't know about that; we have a lot of recordings that indicate you're not really that independent as a journalist.'
After a few seconds of silence, Zucker responds: 'Uh..thank you for your comments', before quickly scrambling to end the discussion quickly.
'So everybody, I think in light of that we'll set up a new system and we'll be back with you. We'll do the rest of the call later,' he said.
The video quickly attracted thousands of replies including one from CNN's Communication division which accused O'Keefe of breaking the law.
'Legal experts say this may be a felony. We've referred it to law enforcement,' the network said.
O'Keefe though shrugged off the threat in a tweet mocking the network saying: 'Our legal experts say Jeff Zucker is just mad and embarrassed. Stay tuned!' 'These statements made by high-up executives at CNN prove that they are simply not interested in being unbiased when reporting on the news.'
The investigation announced on Project Veritas includes many transcripts from the calls in which CNN supposedly 'attacked' President Trump, Fox News host Tucker Carlson, and Republican Senator Lindsey Graham.
'You know, this is what we've come to expect for the last three and a half years, four years, but it clearly is exacerbated by the time that we're in and the issues that he's [Trump] dealing with,' Zucker says.
O'Keefe accused the network of furthering their own political agenda when reporting the news.