Configurable Compartmentation Enables In Vitro Reconstitution of Sustained Synthetic Biology Systems
Configurable Compartmentation Enables In Vitro Reconstitution of Sustained Synthetic Biology Systems
Abstract The compartmentalized and communicative nature of biological cells contributes to the complexity and endurance of living organisms. Current in vitro compartmentalization systems such as droplet emulsions reproduce the compartmentalization property of cells yet fail to recapture the configurability of cellular communication with the environment. To mimic biological cells a step further and expand the capabilities of in vitro compartmentalization, we present here a general strategy that inherits the passive transport phenomenon of biology. The strategy incorporates layered, micrometer-sized, hydrogel-based compartments featuring configurability in composition, functionality, and selective permeability of biomolecules. We demonstrated the unique advantage of our strategy in two scenarios of synthetic biology. First, a compartmentalized cell-free protein synthesis system was reconstituted that could support multiple rounds of reactions. Second, we constructed living bacteria-based biosensors in the hydrogel compartments, which could achieve long-lasting functioning with markedly enhanced fitness in complex environments. Looking forward, our strategy should be widely applicable for constructing complex, robust, and sustained in vitro synthetic molecular and cellular systems, paving the way for their practical applications.
Li Jian、Tian Xintong、Pu Bingchun、Ma Conghui、Xiong Chenwei、Keasling Jay、Li Luyao、Zhang Rong、Zhang Jingwei、Liu Yifan、Ba Fang、Shi Yunfeng、Li Ting
School of Physical Science and Technology, ShanghaiTech UniversitySchool of Physical Science and Technology, ShanghaiTech UniversityDepartment of Immunology and Microbiology, Shanghai Jiao Tong University School of MedicineSchool of Physical Science and Technology, ShanghaiTech UniversitySchool of Physical Science and Technology, ShanghaiTech UniversityJoint BioEnergy Institute, Emeryville||Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory||Department of Chemical and Biomolecular Engineering & Department of Bioengineering, University of CaliforniaSchool of Physical Science and Technology, ShanghaiTech UniversitySchool of Physical Science and Technology, ShanghaiTech UniversityState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan UniversitySchool of Physical Science and Technology, ShanghaiTech UniversitySchool of Physical Science and Technology, ShanghaiTech UniversityState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan UniversityState Key Laboratory of Genetic Engineering, School of Life Sciences, Fudan University
生物工程学分子生物学细胞生物学
Synthetic BiologyCell-free Protein SynthesisBiosensorBiomaterialCompartmentalization
Li Jian,Tian Xintong,Pu Bingchun,Ma Conghui,Xiong Chenwei,Keasling Jay,Li Luyao,Zhang Rong,Zhang Jingwei,Liu Yifan,Ba Fang,Shi Yunfeng,Li Ting.Configurable Compartmentation Enables In Vitro Reconstitution of Sustained Synthetic Biology Systems[EB/OL].(2025-03-28)[2025-08-19].https://www.biorxiv.org/content/10.1101/2022.03.19.484972.点此复制
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