Poly(ethylenimine) as a gene delivery vehicle, and its potential for gene therapy

[摘要] Poly(ethylenimine) (PEI) is a polycation with proven success as a gene delivery vehicle both in vitro and in vivo. The work described herein addresses several aspects of PEI-mediated gene delivery. First, a model system was developed, and an assessment of what PEI molecular weights should be considered for optimal transfection was made. Next, PEI/DNA complexes were examined and their physical characteristics (such as surface charge concentration and buffering capacity) defined in an attempt to further improve PEI/DNA transfection efficiencies. After characterizations were completed, cellular trafficking studies were performed to yield a pictorial overview of the route taken by PEI/DNA complexes during transfection. With the general trafficking route in place, the stage was set for a more in-depth examination of the mechanism behind PEI-mediated transfection. The intracellular route of PEI/DNA complexes during transfection was then re-evaluated with respect to lysosomes, and a novel finding was made that disproves a popular hypothesis that has attempted to explain the success of PEI-mediated transfection. Additionally, the stability of PEI/DNA complexes was established, and applied toward the fact that the complexes are shuttled into cell nuclei during transfection. Finally, the effects of PEI;;s nuclear entry were addressed in terms of secreted cellular products and cell viability.

[发布机构] Rice University

[效力级别] Biochemistry