Background
Macromolecule crowding—and the changes in macromolecule behavior and reactivity observed therein—has been proposed as an important, nonspecific biophysical phenomena with likely evolutionary effects on the organizational functioning of cellular signaling complexes. Recent interest in molecular crowding has led researchers to question the manner in which synthetic cell structures can be used to determine the importance of confinement and volume exclusion on cell behavior. While previous studies involving synthetic cells (or “compartments”) have shed light on macromolecular reactions within liposomes, studies of reaction rates outside the vesicles have yet to be performed. Furthermore, prior generations of synthetic cell reaction vessels were physically larger than naturally-occurring molecular dimensions. As such, more advanced and nuanced materials and assays are needed to fully exploit the potential of synthetic compartments.

Invention Description
The disclosed invention relates to the construction and characterization of synthetic cells that mimic not only the plasma membrane of eukaryotic cells, but also the presence of cytoplasm and internal structures. Operating under the assumption that the macromolecular reactivity, macromolecular crowding, and volume exclusion characteristics of living cells can be replicated and controlled in non-living systems (i.e. synthetic cells), Penn State researchers have methodized the production of two unique vessel compartments: unilamellar liposomes and lithographically-defined depressions in solid substrates. When filled with aqueous solutions containing high concentrations of macromolecules, these synthetic cells can aid in understanding the biomechanics of cellular functions, such as phase separation.

Lithographically–defined compartments offer the advantage of being readily engineered in different shapes and sizes. The ability to manipulate cell shape and volume allows for the direct study of how surface area ratios affect the phase separation behavior of the vessel. Though not as readily derivatized or mass-produced as identical test-structures, membrane-bounded unilamellar liposomes are more accurate models of eukaryotic biology than lithographically-defined compartments. Together, these compartments represent the next generation of synthetic cell technology.

File Number: 2635