Spatially regulated mRNA translation enables functional membrane protein integration in synthetic cells

• 成果类型: Article
• 署名作者: Fu, Hang; Ma, Lijuan; Xu, Chunhua; Li, Jinghua; Ouyang, Haochen; Xie, Congbao; Sun, Yunhai; Tao, Yuanxiao; Wang, Hao; Hu, Shuxin; Fu, Meifang; Zheng, Hai; Zhang, Honghong; Liu, Chenli; Ye, Fangfu; Qiao, Yan; Li, Ming; Lu, Ying
• 署名单位: Chinese Academy of Sciences; Nanjing Institute of Geology & Paleontology, CAS; Institute of Physics, CAS; Chinese Academy of Sciences; University of Chinese Academy of Sciences, CAS; Chinese Academy of Sciences; University of Chinese Academy of Sciences, CAS; Chinese Academy of Sciences; Shenzhen Institute of Advanced Technology, CAS; Chinese Academy of Sciences; Institute of Chemistry, CAS
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.2517323123
• 发表日期: 2026-01-27
• 页码: e2517323123
• 关键词: synthetic; membrane proteins; cell-free protein synthesis; escherichia-coli; vesicles; localization; insertion; DYNAMICS; reveals

摘要

Synthetic cells aim to emulate living systems by reconstituting essential cellular processes within lipid-bound architectures. However, their functional complexity remains constrained by a key challenge: the synthesis and correct integration of hydrophobic membrane proteins via cell-free approaches. Here, inspired by natural cells, we developed a spatially regulated translation strategy in which membrane-anchored mRNAs recruit ribosomes to drive the cotranslational insertion of membrane proteins into lipid bilayers. This design enables efficient in situ synthesis and integration of multiple transmembrane proteins within giant unilamellar vesicles, supporting selective small-molecule transport across membranes. Importantly, the method allows for precise stoichiometric control of membrane protein composition. Together, this work establishes a minimal yet versatile framework for the direct synthesis and integration of membrane proteins, advancing the construction of functional synthetic cells.