An Advanced Application of Protein Microarrays: Cell-Based Assays for Functional Genomics

Abstract

Microarrays have become common tools for approaching different experimental questions: DNA, protein and peptide arrays offer the power of multiplexing the assay and by means of miniaturization technology, the possibility to reduce cost and amount of samples and reagents.

Recently, a novel technology for functional assays has been proposed. Sabatini and co-workers have shown a cell-based microarrays method (1) that relies on the deposition and immobilization of an array of cDNA plasmids on a slide where cells are subsequently plated; the cDNA is then internalized by “reverse transfection” and cells overexpress or downregulate in each single spot the genes of interest. This approach allows the screening of different phenotypes in living cells of many genes in parallel on a single slide.

To overcome some relevant limitations of this approach, we have implemented the technology by means of viral immobilization (2) on a novel surface of cluster-assembled nanostructured TiO₂ (3) previously functionalized with an array of a docking protein.

In this work, we present the detailed development of the “reverse infection cell-microarray based technology” in U2OS cells on a novel coated slide that represents an advanced application of protein arrays.