Precisely Engineered Protein-Based PRINT® Particles for Delivery of Nucleic Acids

Precisely Engineered Protein-Based PRINT® Particles for Delivery of Nucleic Acids Public Deposited

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Abstract

RNA replicon (self-replicating RNA) has long been regarded as an important form of vaccines due to its remarkable immunogenicity and superior safety features. In this study, we report the first non-viral delivery of RNA replicons using protein-based PRINT particles. Cylindrical bovine serum albumin (BSA) particles (diameter (d) 1 micron, height (h) 1 micron) fabricated utilizing PRINT® technology were rendered transiently insoluble using a novel, reductively labile disulfide-based cross-linker. After being cross-linked, the protein particles retain their integrity in aqueous solution and dissolve preferentially under a reducing environment which is found in cytoplasm, the site of action for RNA replicons. Our data reveal that the cross-linker leaves no chemical residue on the amino group it reacts with after being cleaved, which represents a great advantage over traditionally used protein cross-linkers. The particles were loaded with RNA replicons encoding several different proteins i

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