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bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo.

Title: A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo.
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Name(s): Chu, Jun, author
Oh, Younghee, author
Sens, Alex, author
Ataie, Niloufar, author
Dana, Hod, author
Macklin, John J, author
Laviv, Tal, author
Welf, Erik S, author
Dean, Kevin M, author
Zhang, Feijie, author
Kim, Benjamin B, author
Tang, Clement Tran, author
Hu, Michelle, author
Baird, Michelle A, author
Davidson, Michael W, author
Kay, Mark A, author
Fiolka, Reto, author
Yasuda, Ryohei, author
Kim, Douglas S, author
Ng, Ho-Leung, author
Lin, Michael Z, author
Type of Resource: text
Genre: Journal Article
Text
Date Issued: 2016-07-01
Physical Form: computer
online resource
Extent: 1 online resource
Language(s): English
Abstract/Description: Orange-red fluorescent proteins (FPs) are widely used in biomedical research for multiplexed epifluorescence microscopy with GFP-based probes, but their different excitation requirements make multiplexing with new advanced microscopy methods difficult. Separately, orange-red FPs are useful for deep-tissue imaging in mammals owing to the relative tissue transmissibility of orange-red light, but their dependence on illumination limits their sensitivity as reporters in deep tissues. Here we describe CyOFP1, a bright, engineered, orange-red FP that is excitable by cyan light. We show that CyOFP1 enables single-excitation multiplexed imaging with GFP-based probes in single-photon and two-photon microscopy, including time-lapse imaging in light-sheet systems. CyOFP1 also serves as an efficient acceptor for resonance energy transfer from the highly catalytic blue-emitting luciferase NanoLuc. An optimized fusion of CyOFP1 and NanoLuc, called Antares, functions as a highly sensitive bioluminescent reporter in vivo, producing substantially brighter signals from deep tissues than firefly luciferase and other bioluminescent proteins.
Identifier: FSU_pmch_27240196 (IID), 10.1038/nbt.3550 (DOI), PMC4942401 (PMCID), 27240196 (RID), 27240196 (EID), nbt.3550 (PII)
Grant Number: F32 GM117793, P50 GM107615, R01 HL064274, U01 NS090600, R01 GM105404, R01 MH080047
Publication Note: This NIH-funded author manuscript originally appeared in PubMed Central at https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4942401.
Subject(s): Fluorescent Dyes/chemical synthesis
Fluorescent Dyes/pharmacokinetics
Lighting/methods
Luminescent Measurements/methods
Luminescent Proteins/chemical synthesis
Luminescent Proteins/pharmacokinetics
Microscopy, Fluorescence, Multiphoton/methods
Molecular Imaging/methods
Staining and Labeling
Persistent Link to This Record: http://purl.flvc.org/fsu/fd/FSU_pmch_27240196
Host Institution: FSU
Is Part Of: Nature biotechnology.
1546-1696
Issue: iss. 7, vol. 34

Choose the citation style.
Chu, J., Oh, Y., Sens, A., Ataie, N., Dana, H., Macklin, J. J., … Lin, M. Z. (2016). A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo. Nature Biotechnology. Retrieved from http://purl.flvc.org/fsu/fd/FSU_pmch_27240196