A Modular Encapsulation System for Precision Delivery of Proteins, Nucleic Acids, and Small Molecules

Targeted nanoparticles have the potential to revolutionize therapeutics for medical applications. Here, we demonstrate the utility of a flexible precision nanovesicle delivery system for functional delivery of DNA, RNA, proteins, and drugs into target cells. Nanovesicles generated by the membrane sculpting protein caveolin, termed caveospheres, can be loaded with RNA, DNA, proteins, or drugs postsynthesis or incorporate genetically encoded cargo proteins during production without the need for protein purification. Functionalized, fluorescently labeled caveospheres form a modular system that shows high stability in biological fluids and specific uptake by target-positive cells and can deliver proteins, drugs, DNA, and mRNA directly to the cytoplasm and nuclei of only the target cells. The negligible level of off-target transduction and uniform levels of targeted expression demonstrate advantages of the system over lipid-mediated gene delivery. Caveospheres can also be engineered to mimic viral particles by displaying the SARS-CoV-2-RBD protein, enabling the targeted delivery to human bronchial epithelial cells. We demonstrate their application as a targeted transfection system for cells in culture and critically, their efficacy in precision tumor killing <i>in vivo</i>.