Biochemical processes often require spatial regulation and specific microenvironments. The general lack of organelles in bacteria limits the potential of bioengineering complex intracellular reactions. Here we demonstrate Transcriptionally Engineered Addressable RNA Solvent droplets (TEARS) as synthetic microdomains within the Escherichia coli. TEARS are assembled from RNA-binding protein recruitment domains fused to poly-CAG repeats that spontaneously drive liquid-liquid phase separation from the bulk cytoplasm. Targeting TEARS with fluorescent proteins revealed multilayered structures and a non-equilibrium mechanism controlling their composition and reaction robustness. We show that TEARS provide organelle-like bioprocess isolation for sequestering biochemical pathways, controlling metabolic branch points, buffering mRNA translation rates and scaffolding protein-protein interactions. TEARS are a simple and versatile tool for spatially controlling E. coli biochemistry.

中文翻译：

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具有可寻址相分离RNA的大肠杆菌的空间工程

生化过程通常需要空间调节和特定的微环境。细菌中细胞器的普遍缺乏限制了生物工程学复杂细胞内反应的潜力。在这里，我们展示了转录工程可寻址RNA溶剂滴（TEARS）作为大肠杆菌内的合成微区。泪液是从与聚CAG重复序列融合的RNA结合蛋白募集结构域组装而成的，该结构域自发地驱动从大量细胞质中的液相分离。用荧光蛋白靶向TEARS揭示了多层结构和控制其组成和反应稳定性的非平衡机制。我们证明TEARS为隔离生物化学途径，控制代谢分支点，提供细胞器样生物过程隔离，缓冲mRNA的翻译率和脚手架蛋白质间相互作用。TEARS是一种用于空间控制大肠杆菌生物化学的简单而通用的工具。