A Facile and Versatile Platform for Cytosolic Delivery of Proteins in Nanoshells of DNA or RNA: Packaging Options in Multiplexed Delivery

Polyethylenimine (PEI) polymers are used to compact DNA into nanoparticles for delivery into cells. We have shown that PEI-mannose polymers compact DNA into nanoshell-like particles, which can load proteins as well. Here we show that these DNA containers are uniquely versatile for scavenging proteins, irrespective of size, charge, and hydrophobicity from dilute solutions. The number of DNA containers for loading proteins can be controlled independently of the protein loading per container by changing the amounts of DNA and protein in solution. This provides control of the fraction of cells receiving the payload and the relative amounts of DNA and protein per cell. The proteins released inside cells retain enzymatic activity. The proposed technology provides a new way to approach protein delivery by hitchhiking proteins within a facile and well-established DNA-delivery mechanism and by utilizing sugar biophysics to load a wide range of proteins in a single-step process.