Engineering Asymmetric Nanoscale Vesicles for mRNA and Protein Delivery to Cells

Abstract The delivery of therapeutics to cells is crucial for the treatment and prevention of diseases. To enhance targeting and protect therapeutics from degradation, they are often encapsulated in drug delivery vehicles like lipid nanoparticles, lipid vesicles, and viral vectors. However, there is no universal vehicle for all cargo types including small molecules, nucleic acids, and proteins. Here, a method for engineering lipid vesicles with asymmetric leaflets is presented, and their ability to deliver mRNA and proteins to cells is demonstrated. The results show that leaflet asymmetry enhances vesicle uptake by cells, and increases the transfection efficiency with mRNA up to fivefold. Additionally, it is shown that asymmetric vesicles can deliver a variety of proteins. In particular, the delivery of Cas9 proteins and Cas9/sgRNA complexes for gene‐editing is demonstrated. This work expands the design parameters for drug delivery vehicles, enabling more efficient and universal carriers for drug and protein delivery.