Sensor and assay for screening of photosensitizers in living cells
Project ID: D2018-33 Background Photodynamic therapy (PDT) uses light and a photosensitizer in the presence of oxygen to elicit cell death. This technique has been under investigation for use in treatment of cancerMost often used to treat skin cancer, mycosis fungoides, and non-small cell lung cancer. Researchers need to detect the ability of PDTs to disrupt cell signaling in order to develop a cancer treatment. Currently, researchers look at measurement of single oxygen (1O2) or ROS induced oxidation. However, 1O2 and ROS can be the result of other cellular activities. Therefore, there is a need to directly quantify the efficacy of photosensitizers to examine the efficacy of PDT agents in living cells Invention DescriptionResearchers at the University of Toledo have developed an assay that allows for the visualization of PIP2 dislodging from the plasma membrane in living cells. As a research tool, this technology allows for high-throughput screening of photosensitizers, including Rose Bengal, Methylene Blue, FMN, FAD, and Retinal. In the presence of light, singlet oxygen is created by the cells, producing reactive oxygen species, and ultimately causing the peroxidation of PIP2. The sensor reads at wavelengths ~445-594nm, showing solubilization of mcherry tagged PIP2 via fluorescence. ApplicationsScreening for efficacy of photosensitizers and visualization of PIP2 peroxidation in living cells. Advantages• Direct measure rather than measure of by-products• Detects influence photosensitizers on natural biomolecules in cells• Allows for measure of optimum chemical and light doses for PDT
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