RuBisCO is the most abundant enzyme in nature, catalyzing the fixation of CO₂ in photosynthesis. Its common form consists of eight RbcL and eight RbcS subunits, the assembly of which requires a series of chaperones that include RbcX and RuBisCO accumulation factor 1 (Raf1). To understand how these RuBisCO-specific chaperones function during cyanobacterial RbcL₈RbcS₈ (L₈S₈) holoenzyme formation, we solved a 3.3-Å cryo-electron microscopy structure of a 32-subunit RbcL₈Raf1₈RbcX₁₆ (L₈F₈X₁₆) assembly intermediate from Anabaena sp. PCC 7120. Comparison to the previously resolved L₈F₈ and L₈X₁₆ structures together with biochemical assays revealed that the L₈F₈X₁₆ complex forms a rather dynamic structural intermediate, favoring RbcS displacement of Raf1 and RbcX. In vitro assays further demonstrated that both Raf1 and RbcX function to regulate RuBisCO condensate formation by restricting CcmM35 binding to the stably assembled L₈S₈ holoenzymes. Combined with previous findings, we propose a model on how Raf1 and RbcX work in concert to facilitate, and regulate, cyanobacterial RuBisCO assembly as well as disassembly of RuBisCO condensates.