The Beresheet robot is a privately funded venture that aims to land and hop across the lunar surface.
It's a challenging prospect. Only government space agencies from the US, Russia and China have previously managed soft touchdowns.
The 1.5m-high, 585kg Beresheet will begin its mission with a ride to Earth orbit on a SpaceX Falcon 9 rocket from Cape Canaveral in Florida.
Once ejected from this vehicle, the robot will then use its British-built engine to propel itself to the Moon. The journey will take over two months to complete.
SpaceIL, the non-profit behind the project, hopes Beresheet ("In the beginning" in Hebrew) will prove an inspiration to all those who follow its progress.
"By what we’re doing and achieving with the limited resources that we have, and the limited finances we had - I think we showed the Israeli ingenuity," said key donor and SpaceIL chairman, Morris Kahn.
"We show our initiative, and we’ve developed technology, which I think is going to be important. I think we gotta take Israel into space," he told reporters on Monday.
Beresheet grew out of the Google Lunar XPRIZE, which offered financial incentives in 2007 to any non-government-funded team that could pull off a Moon landing.
The lift-off of the Beresheet's Falcon 9 from Cape Canaveral is timed for 20:45 EST, Thursday (01:45 GMT, Friday).
The robot is actually piggybacking a ride on the flight, which has the primary function of placing a new telecoms satellite in orbit for the Indonesian company Pasifik Satelit Nusantara.