Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) catalyses the initial step of photosynthetic CO₂ assimilation, although its catalytic efficiency is very low. Therefore, higher plants must synthesize large amounts of RuBisCO to compensate for its inefficient enzymatic properties. The holoenzyme of RuBisCO consists of 8 large and 8 small subunits, whose genes are individually encoded on two distinct genomes located in the chloroplast and the nucleus, respectively. RuBisCO biosynthesis requires many factors involved in transcription, translation, folding and assembly processes. However, the mechanisms underlying these processes are complex, and therefore the molecular mechanisms and regulation systems of RuBisCO biosynthesis are not yet fully understood. In this review, we introduce recent research on the molecular mechanisms of RuBisCO biosynthesis in higher plants, and discuss future perspectives in this field of research.