Permeability‐Engineered Compartmentalization Enables In Vitro Reconstitution of Sustained Synthetic Biology Systems

作者：Luyao Li, Rong Zhang, Long Chen, Xintong Tian, Ting Li, Bingchun Pu, Conghui Ma, Xiangyang Ji, Fang Ba, Chenwei Xiong, Yunfeng Shi, Xianqiang Mi, Jian Li, Jay D. Keasling, Jingwei Zhang, Yifan Liu · 发表于：Advanced Science · 年份：2022 · DOI：10.1002/advs.202203652 · 被引用次数：27 · 研究领域：Bacteriophages and microbial interactions、Advanced biosensing and bioanalysis techniques、Bacterial Genetics and Biotechnology

Abstract In nature, biological compartments such as cells rely on dynamically controlled permeability for matter exchange and complex cellular activities. Likewise, the ability to engineer compartment permeability is crucial for in vitro systems to gain sustainability, robustness, and complexity. However, rendering in vitro compartments such a capability is challenging. Here, a facile strategy is presented to build permeability‐configurable compartments, and marked advantages of such compartmentalization are shown in reconstituting sustained synthetic biology systems in vitro. Through microfluidics, the strategy produces micrometer‐sized layered microgels whose shell layer serves as a sieving structure for biomolecules and particles. In this configuration, the transport of DNAs, proteins, and bacteriophages across the compartments can be controlled an guided by a physical model. Through permeability engineering, a compartmentalized cell‐free protein synthesis system sustains multicycle protein production; ≈100 000 compartments are repeatedly used in a five‐cycle synthesis, featuring a yield of 2.2 mg mL −1 . Further, the engineered bacteria‐enclosing compartments possess near‐perfect phage resistance and enhanced environmental fitness. In a complex river silt environment, compartmentalized whole‐cell biosensors show maintained activity throughout the 32 h pollutant monitoring. It is anticipated that permeability‐engineered compartmentalization should pave the way for practica...