Basic biophysical studies and pharmacological processes can be investigated by mimicking the intracellular and extracellular environments across an artificial cell membrane construct. The ability to reproduce in vitro simplified scenarios found in live cell membranes in an automated manner has great potential for a variety of synthetic biology and compound screening applications. Here, we present a fully integrated microfluidic system for the production of artificial lipid bilayers based on the miniaturisation of droplet-interface-bilayer (DIB) techniques. The platform uses a microfluidic design that enables the controlled positioning and storage of phospholipid-stabilized water-in-oil droplets, leading successfully to the scalable and automated formation of arrays of DIBs to mimic cell membrane processes. To ensure robustness of operation, we have investigated how lipid concentration, immiscible phase flow velocities and the device geometrical parameters affect the system performance. Finally, we produced proof-of-concept data showing that diffusive transport of molecules and ions across on-chip DIBs can be studied and quantified using fluorescence-based assays.

Phenotypes relevant to oxidative phosphorylation (OXPHOS) in eukaryotes are jointly determined by nuclear and mitochondrial DNA (mtDNA). Thus, in humans, the variable clinical presentations of mitochondrial disease patients bearing the same primary mutation, whether in nuclear or mitochondrial DNA, have been attributed to putative genetic determinants carried in the “other” genome, though their identity and the molecular mechanism(s) by which they might act remain elusive. Here we demonstrate cytoplasmic suppression of the mitochondrial disease-like phenotype of the Drosophila melanogaster nuclear mutant tko25t, which includes developmental delay, seizure sensitivity, and defective male courtship. The tko25t strain carries a mutation in a mitoribosomal protein gene, causing OXPHOS deficiency due to defective intramitochondrial protein synthesis. Phenotypic suppression was associated with increased mtDNA copy number and increased mitochondrial biogenesis, as measured by the expression levels of porin voltage dependent anion channel and Spargel (PGC1α). Ubiquitous overexpression of Spargel in tko25t flies phenocopied the suppressor, identifying it as a key mechanistic target thereof. Suppressor-strain mtDNAs differed from related nonsuppressor strain mtDNAs by several coding-region polymorphisms and by length and sequence variation in the noncoding region (NCR), in which the origin of mtDNA replication is located. Cytoplasm from four of five originally Wolbachia-infected strains showed the same suppressor effect, whereas that from neither of two uninfected strains did so, suggesting that the stress of chronic Wolbachia infection may provide evolutionary selection for improved mitochondrial fitness under metabolic stress. Our findings provide a paradigm for understanding the role of mtDNA genotype in human disease.

Uniformly-sized, nanostructured peptide microparticles are generated by exploiting the ability of enzymes to serve (i) as catalysts, to control self-assembly within monodisperse, surfactant-stabilized water-in-oil microdroplets, and (ii) as destabilizers of emulsion interfaces, to enable facile transfer of the produced microparticles to water. This approach combines the advantages of biocatalytic self-assembly with the compartmentalization properties enabled by droplet microfluidics. Firstly, using microfluidic techniques, precursors of self-assembling peptide derivatives and enzymes are mixed in the microdroplets which upon catalytic conversion undergo molecular self-assembly into peptide particles, depending on the chemical nature of the precursors. Due to their amphiphilic nature, enzymes adsorb at the water-surfactant-oil interface of the droplets, inducing the transfer of peptide microparticles from the oil to the aqueous phase. Ultimately, through washing steps, enzymes can be removed from the microparticles which results in uniformely-sized particles composed of nanostructured aromatic peptide amphiphiles.

Uniformly‐sized, nanostructured peptide microparticles are generated by exploiting the ability of enzymes to serve both as catalysts, to control selfassembly within monodisperse, surfactant‐stabilized waterin‐oil microdroplets, and as destabilizers of emulsion interfaces, to enable facile transfer of the produced microparticles to water. As reported by M. Zagnoni, R. V. Ulijn, and co‐workers on page 285, this approach combines the advantages of biocatalytic self‐assembly with the compartmentalization properties enabled by droplet microfluidics.