Advancing synthetic biology through cell-free protein synthesis

The rapid development of synthetic biology has enabled the production of compounds with revolutionary improvements in biotechnology. DNA manipulation tools have expedited the engineering of cellular sys-tems for this purpose. Nonetheless, the inherent constraints of cellular systems persist, imposing an upper limit on mass and energy conversion efficiencies. Cell-free protein synthesis (CFPS) has demonstrated its potential to overcome these inherent constraints and has been instrumental in the further advancement of synthetic biology. Via the removal of the cell membranes and redundant parts of cells, CFPS has provided flexibility in directly dissecting and manipulating the Central Dogma with rapid feedback. This mini-review summarizes recent achievements of the CFPS technique and its application to a wide range of synthetic biology projects, such as minimal cell assembly, metabolic engineering, and recombinant protein produc-tion for therapeutics, as well as biosensor development for in vitro diagnostics. In addition, current chal-lenges and future perspectives in developing a generalized cell-free synthetic biology are outlined. (c) 2023 The Authors. Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology. This is an open access article under the CC BY license (http://creative-commons.org/licenses/by/4.0/).

Automated summary

The rapid development of synthetic biology has enabled revolutionary improvements in biotechnology. Cell-free protein synthesis (CFPS) has been instrumental in further advancing synthetic biology through its ability to directly manipulate the Central Dogma with flexibility and rapid feedback. This mini-review summarizes recent achievements and applications of CFPS technique in various synthetic biology projects, including minimal cell assembly, metabolic engineering, recombinant protein production for therapeutics, and biosensor development for in vitro diagnostics. It also outlines current challenges and future perspectives in developing a generalized cell-free synthetic biology.