Controlling Secretion in Artificial Cells with a Membrane AND Gate

Abstract

The assembly of channel proteins into vesicle membranes is a useful strategy to control activities of vesicle-based systems. Here, we developed a membrane AND gate that responds to both a fatty acid and a pore-forming channel protein to induce the release of encapsulated cargo. We explored how membrane composition affects the functional assembly of α-hemolysin into phospholipid vesicles as a function of oleic acid content and α-hemolysin concentration. We then showed that we could induce α-hemolysin assembly when we added oleic acid micelles to a specific composition of phospholipid vesicles. Finally, we demonstrated that our membrane AND gate could be coupled to a gene expression system. Our study provides a new method to control the temporal dynamics of vesicle permeability by controlling when the functional assembly of a channel protein into synthetic vesicles occurs. Furthermore, a membrane AND gate that utilizes membrane-associating biomolecules introduces a new way to implement Boolean logic that should complement genetic logic circuits and ultimately enhance the capabilities of artificial cellular systems.

Microfluidic Formation of Honeycomb-Patterned Droplets Bounded by Interface Bilayers via Bimodal Molecular Adsorption

“…When we increased the hemolysin concentration from 62 to 100 nM, we observed no apparent change in FRAP recovery rate. The addition of cholesterol and oleic acid could increase the recovery rate of the FI, as reported previously [ 35 , 36 ]. In addition, this FRAP experiment was particularly challenging because the laser irradiation triggered oil flow, destabilizing the droplets.…”

Barcoding Biological Reactions with DNA‐Functionalized Vesicles

“…Biochemical products synthesized within vesicles as a result of fusion could further expand the functionality of vesicles by generating useful protein products. These protein products could include pore‐forming proteins to release and deliver cargo, or surface antigens that enable sensing of and interactions with the surrounding environment. Ultimately, DNA‐mediated vesicle fusion coupled with the cell‐free synthesis of biological molecules should expand the possible functions of vesicle‐based materials, enabling enhanced sensing, delivery, and biomanufacturing capabilities.…”

Barcoding Biological Reactions with DNA‐Functionalized Vesicles

“…Research Articles deliver cargo, [6,74,75] or surface antigens that enable sensing of and interactions with the surrounding environment. Ultimately,D NA-mediated vesicle fusion coupled with the cellfree synthesis of biological molecules should expand the possible functions of vesicle-based materials,e nabling enhanced sensing,d elivery,a nd biomanufacturing capabilities.…”