Development of a tailorable and tunable mechanism for cell-responsive substrate-mediated gene delivery

Due to the spatial and temporal control as well as the cell-type specificity necessary to extend gene delivery to therapeutic applications, there exists a need to create systems capable of gene transfer that are well-understood and easily manipulated. Furthermore, the creation of such materials will enable further exploration of the correlation between biochemical cues and the resulting cellular responses. In response to this as yet unmet need, a method to promote cell-responsive substrate-mediated gene delivery was developed for this dissertation. Through the use of non-viral gene delivery, flexibility of the vehicle design was incorporated into the system. Using PNA technology, pDNA was able to be specifically tethered to a self-assembled monolayer via an enzymatically-labile peptide tether. This construct was shown to promote cell-responsive delivery while retaining flexibility over the chemical and physical properties of the vehicle and substrate. By alteration of some design parameters including tether number, pDNA surface coverage, and complexation agent, temporal control over the release profile was demonstrated. Furthermore, the ability to extend the applicability of the system was detailed by transitioning to a poly-D-lysine coated substrate upon which the pDNA is immobilized. This dissertation details proof-of-principle work in the formation of a controlled release gene delivery mechanism that may be used to promote understanding of cellular responses to biochemical signaling as well as be extended to use in tissue engineering applications