Description: This project proposes an engineering approach to the creation of a total synthetic cell, underpinning the understanding of assembling life-like systems. In the proposed working plan, we will unblock syntheses from inner cytoplasm, liquid phase separation formed organelles, biomembrane boundary to cell division – a highly functional cellular process. Both the timeliness and relevance of the project are related to its direct usefulness for synthetic biology in which the EU scientific community is significantly involved. The main innovation from this project is coming from the exquisite combination of a precision technology - microfluidics, with protein biology and biophysics. By using microfluidics, monodispersed cell-like constructs will be built at high-throughput.