Single-Cell Biophysics: Measurement, Modulation, and Modeling

Single-Cell Biophysics: Measurement, Modulation, and Modeling

Poster Abstracts

73-POS Board 37 Green Variant of Monomeric Photosensitizing Fluorescent Protein for Photo-Inducible Protein Inactivation and Cell Ablation Yemima D. Riani 1 , Tomoki Matsuda 1,2 , Takeharu Nagai 1,2 . 2 The Institute of Scientific and Industrial Research, Ibaraki, Osaka, Japan. 1 Osaka University, Ibaraki, Osaka, Japan, Photosensitizing fluorescent protein, which generates reactive oxygen species (ROS) upon light irradiation, is known to be useful for protein inactivation and cell ablation. Those give us clues to elucidate protein function, intracellular signaling pathway and intercellular interaction. Red fluorescent protein based monomeric photosensitizer, SuperNova Red (SNR) [1], has been established and overcame the drawbacks of its original dimeric version, KillerRed [2]. Here, we established SuperNova Green (SNG), a green variant of SNR which will be useful to control ROS production spatially, temporally, specifically, and simultaneously when used in combination with SNR. In vitro and in vivo ROS measurement showed that SNG has higher phototoxic activity than the red variant. Specific cell ablation of co-culture HeLa cells expressing SNG and SNR was successfully performed. In conclusion, SNG is useful as a new color variant of photosensitizer and can be used in combination with red variant of SuperNova to perform protein inactivation or cell ablation. References [1] Bulina, M. E., Chudakov, D.M., Britanova, O.V., Yanushevich, Y.G., Staroverov, D.B., Chepurnykh, T.V., Merzlyak, E.M., Shkrob, M.A., Lukyanov, S., Lukyanov, K.A. A genetically encoded photosensitizer. Nat. Biotechnol . 24, 95-99 (2006). [2] Takemoto, K., Matsuda, T., Sakai, N., Fu, D., Noda, M., Uchiyama, S., Kotera, I., Arai, Y., Horiuchi, M., Fukui, K., Ayabe, T., Inagaki, F., Suzuki, H., Nagai, T. SuperNova, a monomeric photosensitizing fluorescent protein for chromophore-assisted light inactivation. Sci. Rep . 3, 2629 (2013).

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