Single step detection of RNA in cells

The overall goal is to develop single step assay in which a single reagent is added to cells and then used directly for the detection of a specific RNA sequence. The main challenge will be to develop a reagent that is compatible with whole blood. This technology will be especially useful for high throughput screening by assaying large libraries of compounds for their effects on the transcription of specific genes. Also, when paired with a small hand-held device, this technology will have strong potential as a point-of-care diagnostic. Small tissue samples from biopsies or blood could be assayed quickly without the up-front RNA isolation step. A variety of compounds and reagents will be screened for their ability to act as a broad-spectrum ribonuclease inhibitor in cell lysates to protect the sample RNA during cell lysis and RNA detection. They will also be evaluated for their affect on the enzymes used in the detection strategy. The overall goal is identify compounds that inhibit ribonuclease activity but do not inhibit enzyme activity and use these compounds in developing the single-step reagent. PROPOSED COMMERCIAL 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. As well, kits will be sold to basic researchers who will value its ease of use and speed.