Condensation of RNA and proteins is central to cellular functions, and the ability to program it would be valuable in synthetic biology and synthetic cell science. Here we introduce a modular platform for engineering synthetic RNA condensates from tailor-made, branched RNA nanostructures that fold and assemble co-transcriptionally. Up to three orthogonal condensates can form simultaneously and selectively accumulate fluorophores through embedded fluorescent light-up aptamers. The RNA condensates can be expressed within synthetic cells to produce membrane-less organelles with a controlled number and relative size, and showing the ability to capture proteins using selective protein-binding aptamers. The affinity between otherwise orthogonal nanostructures can be modulated by introducing dedicated linker constructs, enabling the production of bi-phasic RNA condensates with a prescribed degree of interphase mixing and diverse morphologies. The in situ expression of programmable RNA condensates could underpin the spatial organization of functionalities in both biological and synthetic cells.

RNA 和蛋白质的浓缩是细胞功能的核心，对其进行编程的能力在合成生物学和合成细胞科学中将很有价值。在这里，我们介绍了一个模块化平台，用于从定制的分支 RNA 纳米结构中设计合成 RNA 凝聚物，这些纳米结构折叠和共转录组装。最多可以同时形成三个正交缩合物，并通过嵌入的荧光发光适配体选择性地积累荧光团。RNA 缩合物可以在合成细胞内表达，以产生具有受控数量和相对大小的无膜细胞器，并显示出使用选择性蛋白质结合适配体捕获蛋白质的能力。通过引入专用接头构建体，可以调节其他正交纳米结构之间的亲和力，从而能够产生具有规定程度的界面混合和不同形态的双相 RNA 缩合物。可编程 RNA 凝聚物的原位表达可以支持生物和合成细胞中功能的空间组织。