supernova for cali technique

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SUPERNOVA A MONOMERIC PHOTOSENSITIZING FLUORESCENT PROTEIN FOR CHROMOPHORE-ASSISTED LIGHT INACTIVATION KIWAMU TAKEMOTO ET AL. 2013 SCIENTIFIC REPORTS

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Page 1: Supernova for CALI technique

SUPERNOVA A MONOMERIC PHOTOSENSITIZING FLUORESCENT PROTEIN FOR

CHROMOPHORE-ASSISTED LIGHT INACTIVATION

KIWAMU TAKEMOTO ET AL. 2013SCIENTIFIC REPORTS

Page 2: Supernova for CALI technique

PHOTOSENSITIZERS

Bulina et al., 2006

Page 3: Supernova for CALI technique

CALI CHROMOPHORE-ASSISTED LIGHT INACTIVATION

Photosensitizer ProteinTarget Protein

intense light irradiation Target inactivation

Cellular Death

Page 4: Supernova for CALI technique

PHOTOSENSITIZERS

Li et al., 2012

Page 5: Supernova for CALI technique

KILLER RED VS SUPER NOVA

Takemoto et al., 2013, Bulina et al., 2006

579 nm610 nm

Page 6: Supernova for CALI technique

SuperNova KillerRed EGFP

Cx43

Keratin

Fibrillarin

KILLER RED VS SUPER NOVA

Scale bar 10µmTakemoto et al., 2013,

Page 7: Supernova for CALI technique

EXPERIMENTAL RESULTS

0 15 30

45 60 75 90

Takemoto et al., 2013, Bulina et al., 2006Magnification 100x

Page 8: Supernova for CALI technique

EXPERIMENTAL RESULTSActin-GFP Cofilin-SuperNova

BeforeCALI

AfterCALI

Takemoto et al., 2013Scale bar 10µm

Page 9: Supernova for CALI technique

CALI APPLICATIONS

Metaphase Early Cytokinesis Late Cytokinesis Death Survival

Ou et al., 2012

Page 10: Supernova for CALI technique

CALI IN C.ELEGANS

Scale bar 5µm Scale bar 2,5µm

WT CALI QR.a QR.a (pig-1) QR.p

Page 11: Supernova for CALI technique

REFERENCES

Wei Li, Nico Stuurman, Guangshuo Ou. (2012). Chromophore-assisted laser inactivation in neural development. Neuroscience Bulletin. 28 (4), 333–341.

Maria E Bulina, Konstantin A Lukyanov, Olga V Britanova, Daria Onichtchouk, Sergey Lukyanov & Dmitriy M Chudakov. (2006). Chromophore-assisted light inactivation (CALI) using the phototoxic fluorescent protein KillerRed. Nature Protocols. 1 (2), 947-953.

Maria E Bulina, Dmitriy M Chudakov, Olga V Britanova, Yurii G Yanushevich, Dmitry B Staroverov, Tatyana V Chepurnykh, Ekaterina M Merzlyak, Maria A Shkrob Sergey Lukyanov & Konstantin. (2006). A genetically encoded photosensitizer. Nature Biotechnology. 24 (1), 95-99.

Guangshuo Ou, Nico Stuurman, Michael D’Ambrosio, Ronald D. Vale. (2010). Polarized Myosin Produces Unequal-Size Daughters During Asymmetric Cell Division. Science. 330, 677-679.

Takemoto, K., Matsuda, T., Sakai, N., Fu, D., Noda, M., Uchiyama, S., ... & Nagai, T. (2013). SuperNova, a monomeric photosensitizing fluorescent protein for chromophore-assisted light inactivation. Scientific reports, 3.

Look for diagnosis.com (2014). Chromophore-Assisted Light Inactivation (CALI) using the phototoxic fluorescent protein KillerRed. Available: http://jcs.biologists.org/content/127/8/1621/F1.large.jpg.