Intracellular Delivery of Virus-Like Particles Using a Sheddable Linker

Intracellular targeting is an important aspect of the efficient delivery of drugs and nanotherapeutics. Cytosolic transport of nanomaterials is often an essential requirement for therapeutic delivery into cells but remains a challenge owing to the endosomal trap and eventual lysosomal degradation of cargo. To address this, we designed a functional carrier that escapes the endosome and delivers biological materials into the cell's cytoplasm. For this purpose, we synthesized a glutathione-sensitive linker that connects the well-known mitochondria targeting lipophilic triphenylphosphonium cation (TPP) to the surface of a proteinaceous nanoparticle based on the engineered virus-like particle (VLP) Q|?. Once in the cytosol, the thiol sensitive linker severs the TPP from the nanoparticle, halting its trafficking to the mitochondria, and marooning it in the cytosol. We demonstrate the successful in vitro cytosolic delivery of a VLP loaded with Green Fluorescent Protein, where evenly distributed fluorescence is observed in A549 lung cancer cells after four hours. We further demonstrate successful cytosolic delivery by showing that encapsulating siRNA inside the VLP promotes luminescence silencing in luciferase expressing HeLa cells more efficiently than VLPs that lack our sheddable TPP linker.