Two first-half goals from Omer Atzili secured a famous victory for the hosts to keep their own slim hopes of finishing in the top two of Group H alive with two games to spare.
However, the main focus will be on Juventus as their miserable start to the campaign sunk to new depths with an abject display in Israel.
The result will heap further pressure on the shoulders of Juve boss Massimiliano Allegri, who has now seen his side win just four of their 14 games across all competitions this season.
The Old Lady are now stuck on three points from their four Champions League group games so far, leaving them with a major uphill battle to qualify for the next round in a group that also includes Paris Saint-Germain and Benfica.
Those two teams are joint-top of the group on seven points apiece ahead of their meeting later on Tuesday night, and Juve's defeat means that the winner of that tie in Paris would secure a spot in the last 16.
Maccabi Haifa went into tonight's game on a run of nine consecutive Champions League defeats and looking for their first home win in the competition proper for 20 years.
However, their performance in the reverse tie in Turin will have given them confidence of being able to pick up a result, despite losing that game 3-1, and sure enough this time they made their chances count.
Atzili was the hero of the piece for the Israeli champions, opening the scoring after only seven minutes to give Maccabi Haifa a dream start.
A first-ever victory over the Italian giants then turned from a dream into a reality when Atzili doubled his personal tally shortly before half time, firing the ball into the top corner at the near post to send the home fans wild.
It could have been worse for Juve too, with Wojciech Szczesny denying Frantzdy Pierrot and Atzili, and Tjaronn Chery hitting the crossbar even before that second goal.
The visitors' best moment came on the stroke of half time when Dusan Vlahovic was denied by Josh Cohen, but they offered little when chasing the game in the second half as Allegri's side went down with a whimper once again.
Juve will now go into this weekend's Turin derby on the back of successive high-profile defeats, and with scrutiny over Allegri's job only set to grow after this result.