Abstract

Due to inherent complexity, incompatibility, and variability in living cell systems, biomolecular engineering faces significant obstacles. To find novel solutions to these issues, researchers have turned to cell-free synthetic biology (CFSB), a relatively young field of study. Biochemical processes can be triggered in vitro through cell-free synthesis, providing a wider range of options for biomolecular engineering. Here, we provide a survey of recent advances in cell-free synthesis. These have sparked innovative studies in areas including the synthesis of complex proteins, incorporation of unnatural amino acids, precise post-translational modifications, high-throughput workflow, and synthetic biomolecular network regulation. CFSB has transformed the studies of biological machinery in a profound and practical way for versatile biomolecular engineering applications.

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