Molecular diagnostic using clamping fluorescent probes to amplify RNA detection signal of single-cell assay Problem: The molecular diagnostics market is expected to reach $13 billion by 2024, including cancer detection, blood screening, histology assays, tests for transplant compatibility, and inherited and infectious diseases (The World Market for Molecular Diagnostics, 8th Edition, 2019.) RNA detection is a type of molecular diagnosis that is estimated to make up $800 million by 2024, but it is limited by low signal. Solution: To improve RNA detection, clampFISH physically “clamps” the traditional probes onto the target RNA using common, efficient click-chemistry. The clamped probe attracts many fluorescent amplifier probes, increasing the signal. Its high cost hinders this first clampFISH version. The new clampFISH 2.0 obtains 400-fold signal amplification for detecting RNA while also reducing cost, increasing specificity, and shortening the protocol. Technology: clampFISH 2.0 utilizes redesigned, smaller clamping probes to bind target RNA. The new design is modular, requiring only one custom chemical modification. These improvements reduce the cost of probe production significantly. The new design also improves selectivity, shortening washing steps and overall protocol time. Microscopy images showing a sharper readout with an gene expression example; here the example gene is AXL, which encodes the cell surface receptor tyrosine kinase. Using a faster protocol, clampFISH 2.0 provides a clearer picture (right panel) versus a common alternative for RNA detection (left panel- labelled smFISH for single molecule fluorescence in situ hybridization). Advantages:
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Docket #20-9229