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

H Hang Fu (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics) L Lijuan Ma (Jiangsu Key Laboratory for Biomass-Based Energy and Enzyme Technology, School of Chemistry and Chemical Engineering) C Chunhua Xu (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics) J Jinghua Li (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences) H Haochen Ouyang (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences) C Congbao Xie (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences) Y Yunhai Sun (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences) Y Yuanxiao Tao (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences) H Hao Wang (Division of Quantitative Sciences, Department of Oncology Johns Hopkins University School of Medicine Baltimore Maryland USA) S Shuxin Hu (Beijing National Laboratory for Condensed Matter Physics, Institute of Physics) M Meifang Fu (University of Chinese Academy of Sciences) H Hai Zheng H Honghong Zhang (University of Chinese Academy of Sciences) C Chenli Liu (State Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences) F Fangfu Ye (Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health)) Y Yan Qiao M Ming Li Y Ying Lu

Abstract

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.

Article Details

Volume / Issue Vol. 123, Issue 4
Published January 27, 2026
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (18)

H

Hang Fu

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics

L

Lijuan Ma

Jiangsu Key Laboratory for Biomass-Based Energy and Enzyme Technology, School of Chemistry and Chemical Engineering

C

Chunhua Xu

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics

J

Jinghua Li

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences

H

Haochen Ouyang

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences

C

Congbao Xie

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences

Y

Yunhai Sun

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences

Y

Yuanxiao Tao

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences

H

Hao Wang

Division of Quantitative Sciences, Department of Oncology Johns Hopkins University School of Medicine Baltimore Maryland USA

S

Shuxin Hu

Beijing National Laboratory for Condensed Matter Physics, Institute of Physics

M

Meifang Fu

University of Chinese Academy of Sciences

H

Hai Zheng

H

Honghong Zhang

University of Chinese Academy of Sciences

C

Chenli Liu

State Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences

F

Fangfu Ye

Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health)

Y

Yan Qiao

M

Ming Li

Y

Ying Lu