Water-absorbent polymer networks known as hydrogels are attractive materials from which functional tissue substitutes could be developed using patient-derived stem cells. While hydrogels demonstrate biocompatibility conducive to harboring cells, they lack mechanical and biochemical aspects of the native extracellular matrix (ECM), the intricate microenvironment about which cells function. Conventional photopolymerization-based techniques provide an avenue for incorporating these stimuli within hydrogels, but at the expense of introducing propagating free radicals that are prone to non-specific reactions with biological systems. In light of these limitations, we have developed a strategy for hydrogel formation and modification absent of propagating free radicals, proceeding through oxime ligation moderated by a photocaged alkoxyamine. Upon mild UV light exposure, the photocage is cleaved, liberating the alkoxyamine and permitting localized condensation with an aldehyde to form an oxime-based linkage. After synthesizing multi-arm crosslinkers, functionalized with either benzaldehydes or photocaged alkoxyamines, I demonstrated successful formation of oxime-based hydrogels within minutes of light exposure in the presence of live cells. Through a series of rheological studies, I found polymerization rates and final mechanical properties of these gels could be systematically tuned by varying crosslinker concentrations, light intensity, aniline catalyst concentration, and pH. Harnessing the light-driven aspect of this chemistry, I controlled hydrogel geometry and final mechanical properties by dictating the location and extent of UV exposure, respectively. I then translated photomediated oxime ligation toward the biochemical modification of hydrogels, where full-length proteins containing photocaged alkoxyamines were immobilized in user-defined regions exposed to UV light. The programmability afforded by photomediated oxime ligation can recapitulate mechanical and biochemical stimuli found throughout the ECM, which are dynamic and heterogeneous in their presentation. Consequently, photopolymerized oxime-based hydrogels will enable an enhanced understanding of cell-matrix interactions by serving as improved 3D cell culture platforms, thereby leading to advancements in tissue engineering and regenerative medicine.