Liquid-liquid phase separation has emerged as an important means of intracellular RNA compartmentalization. Some membraneless organelles host two or more compartments serving different putative biochemical roles; the mechanisms for, and functional consequences of, this subcompartmentalization are not yet well understood. Here, we show that adjacent phases of decapeptide-based multiphase model membraneless organelles differ markedly in their interactions with RNA. Additionally, their coexistence introduces new equilibria that alter RNA duplex stability and RNA sorting by hybridization state. These effects require neither biospecific RNA binding sites nor full-length proteins. As such, they are general and point to more primitive versions of mechanisms operating in extant biology that could aid understanding and enable design of functional artificial membraneless organelles.
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