Microcompartmentalized Cell-free Protein Synthesis from Single Molecule Template Dna Using Semi-permeable Alginate Microcapsules

This paper presents a novel microcompartmentalized reaction system for cell-free protein synthesis using semi-permeable microcapsules. We used a microfluidic device to encapsulate template DNA and enzymes for cell-free protein synthesis into semi-permeable microcapsules composed of alginate and polyethylene imine (PEI). The template DNA was encapsulated into individual microcapsules at the concentration less than single molecule per microcapsule by limiting dilution. After encapsulation, the essential substrates for cell-free protein synthesis were fed through the semi-permeable membrane of the microcapsules. The proteins synthesized from the single-molecule template DNA in the microcapsules were successfully detected.