The overall goal is to develop a """"""""Cells-to-Signal"""""""" assay in which a single reagent is added to cells and then used directly for the detection of a specific RNA sequence. The Cells-to-Signal assay will be especially useful by pharmaceutical companies in drug discovery for the high throughput screening of multiple compounds for their effects on the transcription of specific genes. The major challenge will be developing this technology to be compatible with whole blood. Additionally, when paired with a portable device, this technology will have strong potential as a point-of-care diagnostic. In phase I, we demonstrated that this assay was quantitative up to 1000 cells/mu l with cultured cells. In phase II, we will develop complete systems to support the rapid screening of cultured cells in 96- and 384-well formats and to develop assays that are compatible with whole blood. A large component of this research will be to implement methods that prevent genomic DNA detection. Also, various strategies will be tested for their ability to increase the dynamic range of this assay, thereby improving its overall robustness. PROPOSED COMERCIAL APPLICATION: This technology will greatly expedite high throughput screening assays by pharmaceutical companies measuring the effects of compounds on the expression of specific genes. It also has great potential as a point-of-care diagnostics. Kits sold to basic researchers will value its ease of use and speed.
