Surface Immobilization of Adeno-Associated Virus for Localized Gene Delivery

Spatial organization of gene expression is a crucial element in the development of complex native tissues. Tissue engineering strategies continue to expand and overcome significant barriers, but the capacity to achieve spatially controlled gene expression profiles in an engineered construct is still a considerable difficulty. To address this critical need, we have developed a platform technology based on adeno-associated virus (AAV) to enable tissue engineers to obtain specific, spatially organized expression of genes in a population of cells. We have previously shown the ability to localize both cellular adhesion and gene expression using polydimethylsiloxane (PDMS) stamps to pattern various alkanethiol solutions onto gold-coated substrates. We have shown a synergistic effect between the patterning of human fibronectin and adeno-associated virus for the delivery of genes to HeLa cells. Here, we further explore this technology by examining the efficacy of implementing additional adhesive proteins and cell types. We have used quartz crystal microbalance with dissipation monitoring (QCM-D) to analyze adhesive protein and virus attachment. Our flexible platform allows for spatial regulation of a variety of proteins and cell types, making it simple to efficiently deliver upregulation or shRNA cassettes to a target population of cells.