Boyle, William2018-11-282018-11-282017-12http://hdl.handle.net/11299/201073University of Minnesota Ph.D. dissertation. December 2017. Major: Chemistry. Advisor: Theresa Reineke. 1 computer file (PDF); ix, 133 pages.Improved delivery of therapeutic nucleic acid payloads to cells could lead to dramatically improved clinical outcomes for patients suffering from genetic disorders. This work focuses on the use of a trehalose-containing cationic glycopolymer, termed Tr4, to transfect clinically relevant cell types. In particular, the development of gene delivery methods to improve the transfection of cell types associated with the skin disease epidermolysis bullosa are investigated. The sulfated glycosaminoglycan, heparin, is show to form ternary complexes with pDNA and Tr4 leading to dramatically increased transfection efficiency in primary fibroblasts, induced pluripotent stem cells, HepG2, and U87-MG cells. This increase is not caused by improved uptake, but instead appears to be driven by improved intracellular trafficking of polyplexes compared transfection without heparin. Increasing the size of the plasmid cargo from 4.7 kbp to a more therapeutically relevant 10 kbp leads to the complete loss of transfection efficiency in Tr4-heparin transfection of primary fibroblasts and a reduction in transfection efficiency in iPSCs. Co-transfecting with additives meant to increase nuclear localization of the pDNA recovers the efficiency lost by increasing the plasmid size. These techniques allowed for the development of function transfection methods in iPSCs delivering a synthetic transcription activator of collagen type VII. Finally, nanofiber mats containing chondroitin sulfate were developed to scavenge inflammatory molecules from wound exudate.enEpidermolysis BullosaGene DeliveryPolymersTransfectionThesis or Dissertation