Gene expression within a cell population can be directly altered through gene delivery approaches. Traditionally for nonviral delivery, plasmids or siRNA molecules, encoding or targeting the gene of interest, are packaged within nanoparticles. These nanoparticles are then delivered to the media surrounding cells seeded onto tissue culture plastic; this technique is termed bolus delivery. Although bolus delivery is widely utilized to screen for ef fi cient delivery vehicles and to study gene function in vitro, this delivery strategy may not result in ef fi cient gene transfer for all cell types or may not identify those delivery vehicles that will be ef fi cient in vivo. Furthermore, bolus delivery cannot be used in applications where patterning of gene expression is needed. In this chapter, we describe methods that incorporate material surfaces (i.e., surface-mediated delivery) or hydrogel scaffolds (i.e., hydrogel-mediated delivery) to ef fi ciently deliver genes. This chapter includes protocols for surface-mediated DNA delivery focusing on the simplest and most effective methods, which include nonspeci fi c immobilization of DNA complexes (both polymer and lipid vectors) onto serum-coated cell culture polystyrene and self-Assembled monolayers of alkanethiols on gold. Also, protocols for the encapsulation of DNA/cationic polymer nanoparticles into hydrogel scaffolds are described, including methods for the encapsulation of low amounts of DNA (<0.2 g/ L) and high amounts of DNA (>0.2 g/ L) since incorporation of high amounts of DNA poses signi fi cant challenges due to aggregation.