We demonstrate the facile and robust generation of giant peptide vesicles by using an
emulsion transfer method. These robust vesicles can sustain chemical and physical
stresses. The peptide vesicles can host cell-free expression reactions by encapsulating
essential ingredients. We show the incorporation of another cell-free expressed elastin-like
polypeptide into the existing membrane of the peptide vesicles.

Towards creating a suitable compartment for synthetic cells, different cell chassis materials
ranging from phospholipids to polymer to protein-polymer conjugates have been previously
explored. Currently, the majority of cell-like compartments are made of lipid molecules as the
resulting membrane resembles that of a natural cell. However, cell-sized lipid vesicles are
sensitive to physical and chemical stresses and the resulting instability disrupts biochemical
reactions within. Recently, peptide vesicles that are more robust and stable were developed …