title: DNA-based patterning of tethered membrane patches.
abstract: Solid-supported lipid bilayers are useful model systems for mimicking cellular membranes; however, the interaction of the bilayer with the surface can disrupt the function of integral membrane proteins and impede topological transformations such as membrane fusion. As a result, a variety of tethered or cushioned lipid bilayer architectures have been described, which retain the proximity to the surface, enabling surface-sensitive techniques, but physically distance the bilayer from the surface. We have recently developed a method for spatially separating a lipid bilayer from a solid support using DNA lipids. In this system, a DNA strand is covalently attached to a glass slide or SiO2 wafer, and giant unilamellar vesicles (GUVs) displaying the complement rupture to form a planar lipid bilayer tethered above the surface. However, the location of the patch is random, determined by where the DNA-GUV initially binds to its complement. To allow greater versatility and control, we sought a way to pattern tethered membrane patches. We present a method for creating spatially distinct tethered membrane patches on a glass slide using microarray printing. Surface-reactive DNA sequences are spotted onto the slide, incubated to covalently link the DNA to the surface, and DNA-GUVs patches are formed selectively on the printed DNA. By interfacing the bilayers with microfluidic flow cells, materials can be added on top of or fused into the membrane to change the composition of the bilayers. With further development, this approach would enable rapid screening of different patches in protein binding assays and would enable interfacing patches with electrical detectors.
authors.affiliation: Department of Chemistry, Stanford University , Stanford, California 94305-5012, United States.
authors.forename: Laura D
authors.initials: LD
authors.lastname: Hughes
authors.forename: Steven G
authors.initials: SG
authors.lastname: Boxer
mesh_terms: D004247:DNA / Q000737:chemistry*; D008051:Lipid Bilayers / Q000737:chemistry / Q000302:isolation & purification*; D008055:Lipids / Q000737:chemistry*; D015394:Molecular Structure; D010316:Particle Size; D013499:Surface Properties
chemical_list: D008051:Lipid Bilayers; D008055:Lipids; D004247:DNA