Abstract
We developed a novel hybrid cell reactor system via functional fusion of single Escherichia coli protoplast cells, that are deficient in cell wall and expose plasma membrane, with arrayed lipid bilayer chambers on a device in order to incorporate the full set of cytosolic and membrane constituents into the artificial chambers. We investigated gene expression activity to represent the viability of the hybrid cell reactors: over 20% of hybrid cells showed gene expression activity from plasmid or mRNA. This suggests that the hybrid cell reactors retained fundamental activity of genetic information transduction. To expand the applicability of the hybrid cell reactors, we also developed the E. coli-in-E. coli cytoplasm system as an artificial parasitism system. Over 30% of encapsulated E. coli cells exhibited normal cell division, showing that hybrid cells can accommodate and cultivate living cells. This novel artificial cell reactor technology would enable unique approaches for synthetic cell researches such as reconstruction of living cell, artificial parasitism/symbiosis system, or physical simulation to test functionality of synthetic genome.
Highlights
Louis Pasteur proposed the biological dogma, which states that living organisms can only be generated from living organisms, and this dogma has yet to be overturned
We developed a novel method for building an array of hybrid cells from single living bacterial cells and artificial cell reactors displayed on an arrayed lipid bilayer chamber system (ALBiC) device[38]
This was accomplished using the advantageous features of the ALBiC; this device is composed of solid state materials and a representative of soft materials
Summary
Louis Pasteur proposed the biological dogma, which states that living organisms can only be generated from living organisms, and this dogma has yet to be overturned. Full reconstitution of the central reactions constituting the central dogma has become experimentally accessible The reconstitution of such integrated systems in micro-compartments is a step towards full reconstitution of an autonomous self-replication system in vitro. Water-in-oil (W/O) droplets are widely used in artificial cell research because of the high controllability of size and ease of preparation that allows for high-throughput production[24] Because of these advantageous features, quantitative analysis in cell-free assays, such as TX-TL or gene circuits, are performed inside of W/O droplets[2,25,26,27,28]. Pioneering studies have attempted reconstitution of a living cell from cell extract[15,16,33] These studies detected translation activities, the reconstituted artificial cells did not have potentiality for self-replication due to the lack of functional membranes. The cell extract dilution should be minimized to avoid an irreversible loss of viability[36,37]
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