Life takes place thanks to the precise regulation of biological activity in both space and time. Initially attributed only to traditional, membrane-bound organelles, there is now a growing body of evidence that membraneless organelles (MLOs) play a major role in this process. MLOs are biomolecular condensates formed through liquid-liquid phase separation, primarily involving intracellular proteins and RNAs. Bacteria were previously thought to lack the intracellular organization seen in eukaryotic cells, but it is now clear that bacterial processes also involve MLOs. Due to their importance in regulating biological functions, it has become essential to be able to engineer MLOs with defined properties. RNA nanotechnology offers a powerful toolkit for engineering nanoscale structures. Previous work from the group has shown the use of RNA for creating MLOs in vitro. The four-arm RNA nanostructures with terminal kissing loops, termed RNA nanostars, undergo intermolecular self-assembly, leading to the formation of RNA condensates. Our focus is on translating this system into Escherichia coli to create designer RNA MLOs. We developed a method for intracellular production of RNA nanostars and the formation of membraneless compartments inside bacteria. Additionally, we demonstrated the formation of orthogonal, non-mixing RNA compartments. Finally, we showed targeted protein capture within MLOs, paving the way for its application in metabolic engineering and biomanufacturing.
Ng et al. (Sun,) studied this question.