title: Engineering artificial cells by combining HeLa-based cell-free expression and ultrathin double emulsion template.
abstract: Generation of artificial cells provides the bridge needed to cover the gap between studying the complexity of biological processes in whole cells and studying these same processes in an in vitro reconstituted system. Artificial cells are defined as the encapsulation of biologically active material in a biological or synthetic membrane. Here, we describe a robust and general method to produce artificial cells for the purpose of mimicking one or more behaviors of a cell. A microfluidic double emulsion system is used to encapsulate a mammalian cell-free expression system that is able to express membrane proteins into the bilayer or soluble proteins inside the vesicles. The development of a robust platform that allows the assembly of artificial cells is valuable in understanding subcellular functions and emergent behaviors in a more cell-like environment as well as for creating novel signaling pathways to achieve specific cellular behaviors.
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Kenneth K Y
authors.initials: KK
authors.lastname: Ho
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Murray
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI, USA; Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA; Cellular and Molecular Biology Program, University of Michigan, Ann Arbor, MI, USA; Biophysics Program, University of Michigan, Ann Arbor, MI, USA.
authors.forename: Allen P
authors.initials: AP
authors.lastname: Liu
mesh_terms: D058045:Artificial Cells / Q000166:cytology*; D045744:Cell Line, Tumor; D002474:Cell-Free System / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D004655:Emulsions / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008051:Lipid Bilayers / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D044085:Microfluidics / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D004655:Emulsions; D008051:Lipid Bilayers; D008565:Membrane Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D004247:DNA
keywords: Artificial cells; Capillary microfluidics; Cell-free expression; Double emulsion; Lipid bilayer vesicle; Synthetic biology