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Intracellular Protein Delivery using Virus-Like Particles

ID# 2015-4396
Paramyxovirus Packages Foreign Proteins

Technology Summary

Paramyxoviruses naturally bud particles that enclose the viral RNA genomes and transmit infections to new cells. In study of this process, Penn State Inventors defined regions near the C-terminal ends of paramyxovirus nucleocapsid proteins that are sufficient to direct foreign proteins into budding particles. In application of this finding, foreign proteins of interest can now be appended with 15-20 amino acid residue long targeting sequences, resulting in their efficient incorporation into noninfectious virus-like particles (VLPs). The VLPs are natural delivery vehicles, capable of attaching to target mammalian cells and delivering the protein-of-interest directly to the cytoplasm. In proof-of-concept studies, Penn State inventors have demonstrated loading of luciferase, GFP, superoxide dismutase, and Cas9 nuclease into VLPs.

Application & Market Utility

Key advantages of this approach compared to other delivery technologies include: (1) Protein cargos are delivered directly to target cell cytoplasms (not endocytic compartments); (2) Delivers proteins, not nucleic acids, thereby avoiding genetic manipulation of target cells; (3) Short targeting sequences that minimally perturb cargo protein function; (4) Can deliver NLS-bearing cargos to the nuclei of target cells; (5) VLPs accommodate a wide range of cargo sizes ranging from peptides to large proteins such as Cas9 (>1500 aa); and (6) Nearly any cell type can be targeted for delivery, as VLPs bind generically to sialic acid cell surface receptors. Applications may include delivery of DNA-binding transcription factors to induce cellular reprogramming, delivery of Cas9-gRNA complexes to allow genome editing ex vivo, and any situation where direct delivery of a therapeutic protein is desired into cells that are difficult to transfect and/or introduction of foreign nucleic acid sequences to target cells is undesirable.

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