An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging

Dan Yu; William Clay Gustafson; Chun Han; Céline Lafaye; Marjolaine Noirclerc-Savoye; Woo Ping Ge; Desiree A. Thayer; Hai Huang; Thomas B. Kornberg; Antoine Royant; Lily Yeh Jan; Yuh Nung Jan; William A. Weiss; Xiaokun Shu

doi:10.1038/ncomms4626

An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging

Dan Yu, William Clay Gustafson, Chun Han, Céline Lafaye, Marjolaine Noirclerc-Savoye, Woo Ping Ge, Desiree A. Thayer, Hai Huang, Thomas B. Kornberg, Antoine Royant, Lily Yeh Jan, Yuh Nung Jan, William A. Weiss, Xiaokun Shu

Research output: Contribution to journal › Article › peer-review

125 Scopus citations

Abstract

Infrared fluorescent proteins (IFPs) are ideal for in vivo imaging, and monomeric versions of these proteins can be advantageous as protein tags or for sensor development. In contrast to GFP, which requires only molecular oxygen for chromophore maturation, phytochrome-derived IFPs incorporate biliverdin (BV) as the chromophore. However, BV varies in concentration in different cells and organisms. Here we engineered cells to express the haeme oxygenase responsible for BV biosynthesis and a brighter monomeric IFP mutant (IFP2.0). Together, these tools improve the imaging capabilities of IFP2.0 compared with monomeric IFP1.4 and dimeric iRFP. By targeting IFP2.0 to the plasma membrane, we demonstrate robust labelling of neuronal processes in Drosophila larvae. We also show that this strategy improves the sensitivity when imaging brain tumours in whole mice. Our work shows promise in the application of IFPs for protein labelling and in vivo imaging.

Original language	English (US)
Article number	4626
Pages (from-to)	1-7
Number of pages	7
Journal	Nature communications
Volume	5
DOIs	https://doi.org/10.1038/ncomms4626
State	Published - May 15 2014

ASJC Scopus subject areas

General Physics and Astronomy
General Chemistry
General Biochemistry, Genetics and Molecular Biology

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@article{64d8f21db5044c33806f767a44cae51c,

title = "An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging",

abstract = "Infrared fluorescent proteins (IFPs) are ideal for in vivo imaging, and monomeric versions of these proteins can be advantageous as protein tags or for sensor development. In contrast to GFP, which requires only molecular oxygen for chromophore maturation, phytochrome-derived IFPs incorporate biliverdin (BV) as the chromophore. However, BV varies in concentration in different cells and organisms. Here we engineered cells to express the haeme oxygenase responsible for BV biosynthesis and a brighter monomeric IFP mutant (IFP2.0). Together, these tools improve the imaging capabilities of IFP2.0 compared with monomeric IFP1.4 and dimeric iRFP. By targeting IFP2.0 to the plasma membrane, we demonstrate robust labelling of neuronal processes in Drosophila larvae. We also show that this strategy improves the sensitivity when imaging brain tumours in whole mice. Our work shows promise in the application of IFPs for protein labelling and in vivo imaging.",

author = "Dan Yu and Gustafson, {William Clay} and Chun Han and C{\'e}line Lafaye and Marjolaine Noirclerc-Savoye and Ge, {Woo Ping} and Thayer, {Desiree A.} and Hai Huang and Kornberg, {Thomas B.} and Antoine Royant and Jan, {Lily Yeh} and Jan, {Yuh Nung} and Weiss, {William A.} and Xiaokun Shu",

note = "Funding Information: We thank Nathan Joh (Lab of William F. DeGrado) for assistance in gel filtration chromatography. This work was supported by Program for Breakthrough Biomedical Research at UCSF (to X.S.), K08NS079485 and Alex Lemonade Stand Foundation (to W.C.G.), U54CA163155 and the Samuel Waxman Cancer Research Foundation (to W.A.W.), NIH GM030637 (to T.B.K.), the French National Research Agency (Grant ANR-11-JSV5-0009-01 to A.R.). X.S. is an NIH Director{\textquoteright}s New Innovator Award Recipient.",

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doi = "10.1038/ncomms4626",

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AU - Yu, Dan

AU - Gustafson, William Clay

AU - Han, Chun

AU - Lafaye, Céline

AU - Noirclerc-Savoye, Marjolaine

AU - Ge, Woo Ping

AU - Thayer, Desiree A.

AU - Huang, Hai

AU - Kornberg, Thomas B.

AU - Royant, Antoine

AU - Jan, Lily Yeh

AU - Jan, Yuh Nung

AU - Weiss, William A.

AU - Shu, Xiaokun

N1 - Funding Information: We thank Nathan Joh (Lab of William F. DeGrado) for assistance in gel filtration chromatography. This work was supported by Program for Breakthrough Biomedical Research at UCSF (to X.S.), K08NS079485 and Alex Lemonade Stand Foundation (to W.C.G.), U54CA163155 and the Samuel Waxman Cancer Research Foundation (to W.A.W.), NIH GM030637 (to T.B.K.), the French National Research Agency (Grant ANR-11-JSV5-0009-01 to A.R.). X.S. is an NIH Director’s New Innovator Award Recipient.

PY - 2014/5/15

Y1 - 2014/5/15

N2 - Infrared fluorescent proteins (IFPs) are ideal for in vivo imaging, and monomeric versions of these proteins can be advantageous as protein tags or for sensor development. In contrast to GFP, which requires only molecular oxygen for chromophore maturation, phytochrome-derived IFPs incorporate biliverdin (BV) as the chromophore. However, BV varies in concentration in different cells and organisms. Here we engineered cells to express the haeme oxygenase responsible for BV biosynthesis and a brighter monomeric IFP mutant (IFP2.0). Together, these tools improve the imaging capabilities of IFP2.0 compared with monomeric IFP1.4 and dimeric iRFP. By targeting IFP2.0 to the plasma membrane, we demonstrate robust labelling of neuronal processes in Drosophila larvae. We also show that this strategy improves the sensitivity when imaging brain tumours in whole mice. Our work shows promise in the application of IFPs for protein labelling and in vivo imaging.

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An improved monomeric infrared fluorescent protein for neuronal and tumour brain imaging

Abstract

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