Date of Award
Master of Science
Juan C. Noveron
Efficacy of gene therapy is relied upon the improvement of gene delivery vectors into mammalian cells. Although viral-based polynucleotide carriers are still considered as the most common method for gene delivery but the cost effectiveness, nonspecific lethal immune response has evoked the renaissance of non-viral synthetic delivery systems. An initial discovery by Felgnar et al showed Cationic Liposomes (CLs) (having an overall positive charge) when mixed with DNA to form CL-DNA complexes can enhance the transfer of polynucleotides into cells followed by its expression. Presently, a flurry of experimental works is taking place for the better understanding of structural characterization and functional efficacy of supramolecular structures (CLs-DNA complex). This effort has contributed CLs delivery system as the most popular rapidly emerging non-viral method for delivering genes in cells as well as for therapeutic clinical trails.The stability of supramolecular self-assembled structures relies upon pre-organization of molecular receptors effecting molecular recognition and appropriate manipulation of intermolecular non-covalent interaction, hydrogen bonding arrays and ion coordination sites of participating molecular components. Campa et al first reported a novel class of metal mediated amphiphillic molecules that have the ability to generate discrete supramolecular self-assembly in water and condense into CLs-DNA structures capable of transfecting polynucleotides to mammalian cells via redox and ligand exchanged reactions. Here in this work we report a different class of Cu mediated co-ordination amphiphiles which are designed to self organize via co-ordination and hydrogen bonding interactions to cationic metallo liposomes. These CLs are capable of delivering a gene (pEGFPN1) encodes for green fluorescent protein into mammalian cells (HEK 293 T).
We have used transmission electron microscopy (TEM), dynamic light scattering (DLS), and confocal optical microscopy to characterize the structure of CLs-DNA complex and lipoplexes. The transfection efficiency was measured with fluorescent microscopy and fluorescence activated cell sorting (FACS).
Received from ProQuest
Pal, Sarit, "Novel Copper (II) Mediated Transfection Nano-Systems And Biological Properties Of Pyridinium Surfactants" (2013). Open Access Theses & Dissertations. 1902.