The Cajal body (CB) is a conserved non-membrane nuclear structure where several steps of small nuclear RNP particle (snRNP) biogenesis take place. It has been proposed that CB formation follows a liquid–liquid phase separation model, but this hypothesis has never been rigorously tested. Here, we applied live-cell imaging to show that the key CB assembly factor coilin is mobile within the CB, and we revealed a diffusion barrier that limits the coilin exchange between CBs and the nucleoplasm. We generated single aa mutations and demonstrated that RNA-dependent coilin oligomerization and coilin interaction with snRNP are essential for CB formation and maintenance. We applied these data to formulate a mathematical model that links the movement of coilin within the nucleoplasm, CB, and across the boundary with its oligomerization and snRNP binding. Our results illustrate CB as a structure dynamically responding to snRNP assembly and recycling.
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