Permeability-Engineered Compartmentalization Enables In Vitro Reconstitution of Sustained Synthetic Biology Systems.
Permeability-Engineered Compartmentalization Enables In Vitro Reconstitution of Sustained Synthetic Biology Systems.
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渗透性——工程隔室化可实现持续合成生物系统的体外重建
DOI：
10.1002/advs.202203652
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发表时间：
2022-12
期刊：
影响因子：
15.1
通讯作者：
Liu, Yifan
中科院分区：
文献类型：
--
作者：
Li, Luyao;Zhang, Rong;Chen, Long;Tian, Xintong;Li, Ting;Pu, Bingchun;Ma, Conghui;Ji, Xiangyang;Ba, Fang;Xiong, Chenwei;Shi, Yunfeng;Mi, Xianqiang;Li, Jian;Keasling, Jay D.;Zhang, Jingwei;Liu, Yifan
关键词：
在自然界中，生物区室如细胞依赖于动态控制的渗透性进行物质交换和复杂的细胞活动。同样，设计隔室渗透性的能力对于体外系统获得可持续性、稳健性和复杂性至关重要。然而，使体外隔室具有这种能力是具有挑战性的。在这里，提出了一种简单的策略来构建渗透性可配置的隔室，并且这种隔室化的显着优势在体外重建持续的合成生物学系统中显示出来。通过微流体技术，该策略产生了微米级的层状微凝胶，其壳层作为生物分子和颗粒的筛分结构。在这种配置中，DNA、蛋白质和噬菌体跨隔室的运输可以由物理模型控制和引导。通过渗透性工程，一个区室化的无细胞蛋白质合成系统维持了多细胞蛋白质的生产;在五个循环的合成中重复使用了10万个区室，产量为2.2 mg mL−1。此外，工程化的细菌封闭隔室具有近乎完美的噬菌体抗性和增强的环境适应性。在复杂的河流淤泥环境中，分区的全细胞生物传感器在整个32小时污染物监测过程中保持活性。预计渗透性工程化区室化将为实际的合成生物学应用铺平道路，例如绿色生物生产，环境传感和基于细菌的治疗。提出了一种渗透性工程化分区策略，以实现持续和稳健的合成生物学系统的设计和构建。基于水凝胶的核壳隔室通过高通量微流体制造，并通过渗透性调节显示出对生物分子和颗粒运输的可控性。该策略导致可回收的无细胞蛋白质合成和具有增强的环境适应性的区室化全细胞生物传感器。
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 practical synthetic biology applications such as green bioproduction, environmental sensing, and bacteria‐based therapeutics. A permeability‐engineerable compartmentalization strategy is presented to enable the design and construction of sustained and robust synthetic biology systems. Hydrogel‐based core–shell compartments are fabricated via high‐throughput microfluidics and show controllability over the transport of biomolecules and particles through permeability adjustment. The strategy leads to recyclable cell‐free protein synthesis and compartmentalized whole‐cell biosensors that feature enhanced environmental fitness.
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影响因子：
2.4
作者：
Abate, Adam R.;Rotem, Assaf;Weitz, David A.
通讯作者：
Weitz, David A.
影响因子：
1.8
作者：
Hsu, Hsiao-Ping;Paul, Wolfgang;Binder, Kurt
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影响因子：
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作者：
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DOI：
10.1073/pnas.1321321111
发表时间：
2014-04-01
影响因子：
11.1
作者：
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通讯作者：
Silver, Pamela A.
影响因子：
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作者：
Mann, Stephen
通讯作者：
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