A magnetic tunnel junction (MTJ) is the core component in memory technologies, such as the magnetic random-access memory, magnetic sensors and programmable logic devices. In particular, MTJs based on two-dimensional van der Waals (vdW) heterostructures offer unprecedented opportunities for low power consumption and miniaturization of spintronic devices. However, their operation at room temperature remains a challenge. Here, we report a large tunnel magnetoresistance (TMR) of up to 85% at room temperature ( T = 300 K) in vdW MTJs based on a thin (< 10 nm) semiconductor spacer WSe 2 layer embedded between two Fe 3 GaTe 2 electrodes with intrinsic above-room-temperature ferromagnetism. The TMR in the MTJ increases with decreasing temperature up to 164% at T = 10 K. The demonstration of TMR in ultra-thin MTJs at room temperature opens a realistic and promising route for next-generation spintronic applications beyond the current state of the art.