 |
Example of differential activation of interrelated genetic promoter-
reporters by various polymers. Results shown are normalized for
each promoter-reporter to tissue culture polystyrene (TCPS).
|
All cell actions (e.g. differentiation, proliferation, inflammation, apoptosis) require genetic upregulation of target genes to carry out cellular functions, and many genes in these pathways are known to be affected by the interactions of the cells with their growth matrix substrate. We anticipate that an understanding of the gene regulation response to different polymer substrates within and across polymer libraries will guide our polymer chemists to design the optimal polymer for the desired application. Using high-throughput analyses of biomaterial-nfluenced gene regulation, this research seeks to identify biomaterials that upregulate genes leading to bone formation, vascularization of newly formed tissue, and enhancment of nerve regeneration.
The Kohn Lab has the capability to synthesize a large array of different polymers via combinatorial synthesis. Accordingly, our methodology for pinpointing materials that promote specific genetic outcomes can be used for large numbers of polymers as well as for studying the effects of a single test substrate.
Our approach involves three preparatory steps: (1) We select a cassette of genes whose activation is indicative of our desired cellular outcome; (2) We genetically fuse the promoter regions of these genes with reporter genes (such as luciferase or fluorescent proteins); (3) We virally transduce the promoter-reporter constructs into the cell of interest. Cells virally transduced to contain multiple promoter-reporters are then seeded on polymer libraries and assayed for gene activation. The presence of multiple promoter-reporters is essential to provide a time course for gene activation and to eliminate false positives.
Project Leader: Jared Bushman, PhD
Funding Source: National Institutes of Health (RESBIO, EB001046)
Our Science