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Single-Cell Biophysics: Measurement, Modulation, and Modeling
Poster Abstracts
81
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).