Optimization and Application of the Purified Cell-Free System.

Jun Ni, Jiawei Wang, Chaofeng Li, Chao Liao, Hengrun Li, Haotian Zheng, Liangxu Liu

Journal: Chembiochem : a European journal of chemical biology 2025;26(11):e202500176

PMID: 40181437

Abstract

Cell-free protein synthesis (CFPS) is extensively applied in biotechnological research. Unlike the lysate-based CFPS system, the alternative purified cell-free system, protein synthesis using recombinant elements (PURE), has garnered a great attention due to its controllability and flexibility of adjusting individual component for in vitro transcription and translation research. The PURE system has been widely studied in the synthetic biological field, and optimized for its application in prototyping research, non-canonical amino acid incorporation and synthetic cell construction. Herein, the recent optimization strategies for a simple and enhanced PURE system are discussed. In addition, the recent application via the PURE system and the current challenges for the development of a more robust and controllable PURE are highlighted.

Address: State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.; Zhangjiang Institute for Advanced Study, Shanghai Jiao Tong University, Shanghai, 200240, China.

DOI: 10.1002/cbic.202500176

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https://doi.org/10.1002/cbic.202500176

MeSH Terms: Cell-Free System, Protein Biosynthesis, Recombinant Proteins