Molecular and Cellular Bioengineering Research Experiences for Undergraduates (REU) Program
SINGLE MOLECULE STUDY OF DNA PACKAGING BY His-Lys GENE DELIVERY PEPTIDES
Histidine-lysine (HK) polymers with specific sequence and branching structures showed great promise for gene delivery in vitro and in vivo. Since HK polymers are synthesized by solid phase synthesis, tailored modifications of their sequence and branching can be employed at single amino acid residue level. This ability to control molecular structure systematically makes the HK polymers ideal candidates for studying molecular mechanism of DNA condensation and release using single molecule technique. In this project, we will investigate the molecular processes of DNA condensation and release by HK polymers at the single-molecule level using optical tweezers. We will directly measure kinetics of HK polymer:plasmid DNA complex (polyplex) formation, the compactness of polyplex, and the kinetics of DNA release from the complex under various ionic conditions in a real time. We will elucidate how systemic structural variations of HK polymers affect DNA condensation and release, identifying key structural features of HK polymer that control these processes. We anticipate that detailed mechanistic understanding of the HK polymers at molecular level will enhance further development of HK polymer and other non-viral carriers with improved transfection efficiency.
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