Holistic engineering of cell-free systems through proteome-reprogramming synthetic circuits
Holistic engineering of cell-free systems through proteome-reprogramming synthetic circuits
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DOI：
10.1038/s41467-020-16900-7
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发表时间：
2020-06-19
影响因子：
16.6
通讯作者：
Tan, Cheemeng
中科院分区：
文献类型：
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作者：
Contreras-Llano, Luis E.;Meyer, Conary;Tan, Cheemeng
合成生物学一直专注于从宿主细胞垂直操作的工程遗传模块。然而，可以设计一个合成生物模块来重新编程宿主蛋白质组，这反过来又增强了合成模块的功能。在这里，我们将这种整体合成生物学的概念应用到无细胞系统的工程中，利用细胞内代谢网络之间的串扰，导致更有利于蛋白质合成的蛋白质环境。具体地说，我们证明了表达翻译机制的局部模块可以重新编程细菌蛋白质组，改变700多种蛋白质的表达水平。由此产生的反馈产生了一个无细胞系统，该系统可以合成荧光记者、蛋白质纳米笼和基因编辑核酸酶Cas9，其表达水平比传统的无细胞系统高5倍。我们的工作展示了一种整合合成和系统生物学概念的整体方法，以实现仅通过局部、正交电路无法实现的结果。合成的生物模块可以用来重新编程宿主蛋白质组，这反过来又增强了合成模块的功能。作者使用这种整体合成生物学的方法来为无细胞蛋白质合成创造更有利的环境。
Synthetic biology has focused on engineering genetic modules that operate orthogonally from the host cells. A synthetic biological module, however, can be designed to reprogram the host proteome, which in turn enhances the function of the synthetic module. Here, we apply this holistic synthetic biology concept to the engineering of cell-free systems by exploiting the crosstalk between metabolic networks in cells, leading to a protein environment more favorable for protein synthesis. Specifically, we show that local modules expressing translation machinery can reprogram the bacterial proteome, changing the expression levels of more than 700 proteins. The resultant feedback generates a cell-free system that can synthesize fluorescent reporters, protein nanocages, and the gene-editing nuclease Cas9, with up to 5-fold higher expression level than classical cell-free systems. Our work demonstrates a holistic approach that integrates synthetic and systems biology concepts to achieve outcomes not possible by only local, orthogonal circuits. Synthetic biological modules can be used to reprogram host proteomes, which in turn enhance the function of the synthetic modules. The authors use this holistic synthetic biology approach to engineer a more favorable environment for cell-free protein synthesis.