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Open Access Dissertation
Doctor of Philosophy (PhD)
Year Degree Awarded
Month Degree Awarded
Vincent M Rotello
Biomedical Engineering and Bioengineering | Biotechnology | Cell Biology | Materials Chemistry
Direct cytoplasmic delivery of gene editing nucleases such CRISPR/Cas9 systems and therapeutic proteins provides enormous opportunities in curing human genetic diseases, and assist research in basic cell biology. One approach to attain such a goal is through engineering nanotechnological tools to mimic naturally existing intra- and extracellular protein delivery/transport systems. Nature builds transport systems for proteins and other biomolecules through evolution-derived sophisticated molecular engineering. Inspired by such natural assemblies, I employed molecular engineering approaches to fabricate self-assembled nanostructures to use as intracellular protein delivery tools. Briefly, proteins and gold nanoparticles were co-engineered to carry complementary electrostatic recognition elements. When these materials were mixed together, they formed highly sophisticated, multi-layered, and hierarchical self-assembled nanostructures of few hundred-nanometer size. These structures carried a large number of engineered proteins, got fused to cell membrane upon incubation, and delivered the encapsulated protein content directly into cell cytoplasm. Using this technology, we delivered a wide range of proteins, and CRISPR/Cas9-ribonucleoprotein that resulted high efficient gene editing.
Mout, Rubul, "PROTEIN-NANOPARTICLE CO-ENGINEERING: SELF-ASSEMBLY, INTRACELLULAR PROTEIN DELIVERY, AND CRISPR/CAS9-BASED GENE EDITING" (2017). Doctoral Dissertations. 1112.