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Near-infrared fluorescent probes based on bacterial phytochromes for in vivo imaging

Cel

Non-invasive monitoring of deep-tissue developmental, metabolic and pathogenic processes will advance modern biology. Imaging of live mammals using fluorescent probes is more feasible within the near-infrared (NIR) transparency window (NIRW: 650-900 nm) where hemoglobin and melanin absorbance significantly decreases, and water absorbance is still low. The most red-shifted fluorescent proteins (FPs) of the GFP-like family exhibit fluorescence outside of the NIRW and suffer from low brightness and modest photostability. Natural bacterial phytochrome photoreceptors (BphPs) utilize low molecular weight biliverdin as a chromophore and provide many advantages over other chromophore binding proteins. First, unlike the chromophores of non-bacterial phytochromes, biliverdin is ubiquitous in mammals. This makes BphP applications in mammalian cells, tissues and mammals as easy as conventional GFP-like FPs, without supplying chromophore through an external solution. Second, BphPs exhibit NIR absorbance and fluorescence, which are red-shifted relative to that of any other phytochromes, and lie within the NIRW. This makes BphPs spectrally complementary to GFP-like FPs and available optogenetic tools. Third, independent domain architecture and conformational changes upon biliverdin photoisomerization make BphPs attractive templates to design various photoactivatable probes. Based on the analysis of the photochemistry and structural changes of BphPs we plan to develop three new types of the BphP-based probes. These include bright and spectrally resolvable permanently fluorescent NIRFPs, NIRFPs photoswitchable either irreversibly or repeatedly with non-phototoxic NIR light, and NIR reporters and biosensors. The resulting NIR probes will extend fluorescence imaging methods to deep-tissue in vivo macroscopy including multicolor cell and tissue labeling, cell photoactivation and tracking, detection of enzymatic activities and protein interactions in mammalian tissues and whole animals.

Zaproszenie do składania wniosków

ERC-2013-ADG
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System finansowania

ERC-AG - ERC Advanced Grant

Instytucja przyjmująca

HELSINGIN YLIOPISTO
Wkład UE
€ 2 496 946,00
Adres
YLIOPISTONKATU 3
00014 Helsingin Yliopisto
Finlandia

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Region
Manner-Suomi Helsinki-Uusimaa Helsinki-Uusimaa
Rodzaj działalności
Higher or Secondary Education Establishments
Kierownik naukowy
Vladislav Verkhusha (Prof.)
Kontakt administracyjny
Kaisa Ainala (Ms.)
Linki
Koszt całkowity
Brak danych

Beneficjenci (1)