Protein Expression inside Oil-free Giant Vesicles by Using Pulsed Jet Flow Method

This paper describes protein expression inside highly stable and oil-free giant vesicles (GVs) that are prepared by deforming a planar lipid membrane using a pulsed jet flow (Fig. 1). We found that the encapsulation efficiency of plasmid DNA into GVs formed by the pulsed jet flow was 7 times higher than that by conventional methods (e.g. gentle hydration method). Moreover, green fluorescent protein (GFP) was successfully expressed in GVs when GVs initially contained plasmid DNA and a cell-free protein synthesis solution. We believe that this method will be useful for the rapid and efficient encapsulation of highly-dense biological molecules, which is essential toward the production of minimal cells.