Higher-order structures that originate from the specific and reversible DNA-directed self-assembly of microscopic building blocks hold great promise for future technologies. Here, we functionalized biotinylated soft colloid oil-in-water emulsion droplets with biotinylated single-stranded DNA oligonucleotides using streptavidin as an intermediary linker. We show the components of this modular linking system to be stable and to induce sequencespecific aggregation of binary mixtures of emulsion droplets. Three length scales were thereby involved: nanoscale DNA base pairing linking microscopic building blocks resulted in macroscopic aggregates visible to the naked eye. The aggregation process was reversible by changing the temperature and electrolyte concentration and by the addition of competing oligonucleotides. The system was reset and reused by subsequent refunctionalization of the emulsion droplets. DNA-directed self-assembly of oil-in-water emulsion droplets, therefore, offers a solid basis for programmable and recyclable soft materials that undergo structural rearrangements on demand and that range in application from information technology to medicine.C omplex higher-order structures that spontaneously arise from the specific and reversible self-assembly of simple building blocks hold great promise to become the foundation of tomorrow's technology (1). In the context of DNA-directed self-assembly of hard colloids and artificial vesicles, linear DNA oligonucleotides were shown to be adhesive elements that allowed the self-assembly process not only to be specific but also to be reversible by a variety of external stimuli such as temperature (2-4), changes in the electrolyte concentration (4, 5), addition of competing oligonucleotides (6), and exploitation of the secondary structure of DNA oligonucleotides (7).Oil-in-water (o/w) emulsion droplets (EDs) offer a robust, configurable, and recyclable soft-colloid system with interesting properties for the fabrication of new delivery systems and advanced synthetic materials with adaptable properties (8). By exploiting the well-characterized ligand-receptor pair biotin-streptavidin, EDs were immobilized on glass surfaces (9) and actively transported along microtubules (10). In the context of self-assembly, both polymer-mediated (11-13) and biotin-streptavidin-mediated aggregation of EDs was shown (14). Despite the advantages a combination of EDs as building blocks and DNA oligonucleotides as linking agent offers, a protocol for DNA-directed self-assembly of EDs is currently missing.To close this gap, here we prepared an o/w emulsion composed of heavier-than-water diethyl phthalate (DEP) dispersed in an aqueous medium, stabilized by phospholipids, and functionalized with single-stranded DNA (ssDNA) oligonucleotides. Palmitoyloleoyl phosphatidylcholine (POPC) and distearoylphosphatidyl ethanolamine (DSPE) in a 9:1 molar ratio were used to stabilize the EDs. The DSPE was grafted with flexible poly(ethylene glycol) (PEG) tethers with a molecular weight of 2,000 (DSPE-PEG...
The technological aspects of artificial vesicles as prominent cell mimics are evolving toward higher-order assemblies of functional vesicles with tissuelike architectures. Here, we demonstrate the spatially controlled DNA-directed bottom-up synthesis of complex microassemblies and macroassemblies of giant unilamellar vesicles functionalized with a basic cellular machinery to express green fluorescent protein and specified neighbor-to-neighbor interactions. We show both that the local and programmable DNA pairing rules on the nanoscale are able to direct the microscale vesicles into macroscale soft matter assemblies and that the highly sensitive gene-expression machinery remains intact and active during multiple experimental steps. An in silico model recapitulates the experiments performed in vitro and covers additional experimental setups highlighting the parameters that control the DNA-directed bottom-up synthesis of higher-order self-assembled structures. The controlled assembly of a functional vesicle matrix may be useful not only as simplified natural tissue mimics but also as artificial scaffolds that could interact and support living cells.
Vesicles and other bilayered membranous structures can self-assemble from single hydrocarbon chain amphiphiles. Their formation and stability are highly dependent upon experimental conditions such as ionic strength, pH and temperature. The addition of divalent cations, for example, often results in the disruption of vesicles made of a single fatty acid species through amphiphile precipitation. However, membranes composed of amphiphile mixtures have been shown to be more resistant to low millimolar concentrations of divalent cations at room temperature. In this report, several mixtures of amphiphiles are examined for their propensity to self-assemble into membranous vesicular structures under extreme environmental conditions of low pH, high ionic strengths and temperatures. In particular, mixtures of decylamine with polar co-surfactants were found to efficiently form membranes under these conditions far away from those normally supporting vesicle formation. We further examined decanoic acid/decylamine mixtures in detail: at pH 2 in low ionic strength solutions, the amphiphiles formed oily or crystalline structures, however the introduction of salts or/and strong acids in conjunction with high temperature induced a stable vesiculation. Thus, extreme environments, such as volcanic or vent environments, whose environmental conditions are known to support high chemical reactivity, could have harbored and most significantly promoted the formation of simple organic compartments that preceded cells.
Abstract:An all-polymer photonic crystal slab sensor is presented, and shown to exhibit narrow resonant reflection with a FWHM of less than 1 nm and a sensitivity of 31 nm/RIU when sensing media with refractive indices around that of water. This results in a detection limit of 4.5 × 10 −6 RIU when measured in conjunction with a spectrometer of 12 pm/pixel resolution. The device is a two-layer structure, composed of a low refractive index polymer with a periodically modulated surface height, covered with a smooth upper-surface high refractive index inorganic-organic hybrid polymer modified with ZrO 2 -based nanoparticles. Furthermore, it is fabricated using inexpensive vacuum-less techniques involving only UV nanoreplication and polymer spin-casting, and is thus well suited for single-use biological and refractive index sensing applications.
The ability to manipulate and fuse nano-compartmentalized volumes addresses a demand for spatiotemporal control in the field of synthetic biology, for example in the bottom-up construction of (bio)chemical nanoreactors and for the interrogation of enzymatic reactions in confined space. Herein, we mix entrapped sub-attoliter volumes of liposomes (~135 nm diameter) via lipid bilayer fusion, facilitated by the hybridization of membrane-anchored lipidated oligonucleotides. We report on an improved synthesis of the membrane-anchor phosphoramidites that allows for a flexible choice of lipophilic moiety. Lipid-nucleic acid conjugates (LiNAs) with and without triethylene glycol spacers between anchor and the 17 nt binding sequence were synthesized and their fusogenic potential evaluated. A fluorescence-based content mixing assay was employed for kinetic monitoring of fusion of the bulk liposome populations at different temperatures. Data obtained at 50 °C indicated a quantitative conversion of the limiting liposome population into fused liposomes and an unprecedently high initial fusion rate was observed. For most conditions and designs only low leakage during fusion was observed. These results consolidate LiNA-mediated membrane fusion as a robust platform for programming compartmentalized chemical and enzymatic reactions.
Label-free, real-time detection of concentration gradients is demonstrated in a microfluidic H-filter, using an integrated photonic crystal slab sensor to monitor sample refractive index with spatial resolution. The recorded diffusion profiles reveal root-mean-square diffusion lengths for non-fluorescing and non-absorbing molecules, both small (glucose, 180 Da) and large (bovine serum albumin, 67 kDa).
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.