法国巴黎大学Ariel B. Lindner、Haotian Guo等研究人员合作利用相分离的RNA对大肠杆菌进行空间工程设计。相关论文发表在2022年9月29日出版的《细胞》杂志上。

研究人员展示了大肠杆菌中的合成无膜细胞器，称为转录工程可寻址RNA溶剂滴（TEARS）。TEARS是由RNA结合蛋白招募结构域融合到聚CAG重复序列中组装而成的，它能自发地驱动液-液相分离，与大量细胞质分离。用荧光蛋白靶向TEARS显示了多层结构，其组成和反应的稳健性受非平衡动力学的制约。研究人员表明，TEARS提供了类似细胞器的生物过程隔离，用于封存生化途径，控制代谢分支点，缓冲mRNA翻译率，以及构建蛋白质-蛋白质相互作用的支架。研究人员预计TEARS将成为空间控制大肠杆菌生物化学的一个简单而通用的工具。特别是，TEARS的模块化设计使得应用不需要对表达进行微调，从而简化了生物工程的设计-建造-测试周期。

据介绍，生物化学过程往往需要空间调节和特定的微环境。细菌中普遍缺乏细胞器，这限制了对复杂的细胞内反应进行生物工程的潜力。

附：英文原文

Title: Spatial engineering of E. coli with addressable phase-separated RNAs

Author: Haotian Guo, Joseph C. Ryan, Xiaohu Song, Adeline Mallet, Mengmeng Zhang, Victor Pabst, Antoine L. Decrulle, Paulina Ejsmont, Edwin H. Wintermute, Ariel B. Lindner

Issue&Volume: 2022/09/29

Abstract: Biochemical processes often require spatial regulation and specific microenvironments.The general lack of organelles in bacteria limits the potential of bioengineeringcomplex intracellular reactions. Here, we demonstrate synthetic membraneless organellesin Escherichia coli termed transcriptionally engineered addressable RNA solvent droplets (TEARS). TEARSare assembled from RNA-binding protein recruiting domains fused to poly-CAG repeatsthat spontaneously drive liquid-liquid phase separation from the bulk cytoplasm. TargetingTEARS with fluorescent proteins revealed multilayered structures with compositionand reaction robustness governed by non-equilibrium dynamics. We show that TEARS provideorganelle-like bioprocess isolation for sequestering biochemical pathways, controllingmetabolic branch points, buffering mRNA translation rates, and scaffolding protein-proteininteractions. We anticipate TEARS to be a simple and versatile tool for spatiallycontrolling E. coli biochemistry. Particularly, the modular design of TEARS enables applications withoutexpression fine-tuning, simplifying the design-build-test cycle of bioengineering.

DOI: 10.1016/j.cell.2022.09.016

Source: https://www.cell.com/cell/fulltext/S0092-8674(22)01174-6