Summary of 217-Sakai-iRFPactivity

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Title
Time-series images of iRFP actitivy in cultured fission or budding yeast cells with BV or PCB.
Description
-
Relase date
2022-03-31
Updated date
-
License
CC-BY
Kind
Image data based on Experiment
Number of Datasets
8 ( Image datasets: 8, Quantitative data datasets: 0 )
Size of Datasets
437.0 MB ( Image datasets: 437.0 MB, Quantitative data datasets: 0 bytes )

Organism(s)
Schizosaccharomyces pombe, Saccharomyces cerevisiae
Strain(s)
SK482, SK483, SK276
Gene symbol(s)
hmx1

Datatype
-
Molecular Function (MF)
biliverdin reductase (NAD(P)+) activity
Biological Process (BP)
phycocyanobilin biosynthetic process
Cellular Component (CC)
-
Biological Imaging Method
confocal microscopy
X scale
-
Y scale
-
Z scale
-
T scale
-

Image Acquisition
Experiment type
-
Microscope type
-
Acquisition mode
-
Contrast method
-
Microscope model
-
Detector model
-
Objective model
-
Filter set
-

Related paper(s)

Keiichiro Sakai, Yohei Kondo, Hiroyoshi Fujioka, Mako Kamiya, Kazuhiro Aoki, Yuhei Goto (2021) Near-infrared imaging in fission yeast using a genetically encoded phycocyanobilin biosynthesis system., Journal of cell science, Volume 134, Number 24

Published in 2021 Dec 15 (Electronic publication in Dec. 16, 2021, midnight )

(Abstract) Near-infrared fluorescent protein (iRFP) is a bright and stable fluorescent protein with near-infrared excitation and emission maxima. Unlike the other conventional fluorescent proteins, iRFP requires biliverdin (BV) as a chromophore. Here, we report that phycocyanobilin (PCB) functions as a brighter chromophore for iRFP than BV, and that biosynthesis of PCB allows live-cell imaging with iRFP in the fission yeast Schizosaccharomyces pombe. We initially found that fission yeast cells did not produce BV and therefore did not show any iRFP fluorescence. The brightness of iRFP-PCB was higher than that of iRFP-BV both in vitro and in fission yeast. We introduced SynPCB2.1, a PCB biosynthesis system, into fission yeast, resulting in the brightest iRFP fluorescence. To make iRFP readily available in fission yeast, we developed an endogenous gene tagging system with iRFP and all-in-one integration plasmids carrying the iRFP-fused marker proteins together with SynPCB2.1. These tools not only enable the easy use of multiplexed live-cell imaging in fission yeast with a broader color palette, but also open the door to new opportunities for near-infrared fluorescence imaging in a wider range of living organisms. This article has an associated First Person interview with the first author of the paper.
(MeSH Terms)

Contact
Kazuhiro Aoki , Yuhei Goto , National Institutes of Natural Sciences , National Institutes of Natural Sciences , Exploratory Research Center on Life and Living Systems (ExCELLS) , Exploratory Research Center on Life and Living Systems (ExCELLS) , Quantitative Biology Research Group , Quantitative Biology Research Group
Contributors


Dataset List of 217-Sakai-iRFPactivity

#
Dataset ID
Kind
Size
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SSBD:OMERO
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# 7566
Datast ID Fig1C_BV_500uM
Dataset Kind Image data
Dataset Size 91.5 MB
4D view
SSBD:OMERO
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# 7567
Datast ID Fig1D_BV_500uM
Dataset Kind Image data
Dataset Size 35.6 MB
4D view
SSBD:OMERO
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# 7568
Datast ID Fig3D_BV_125uM
Dataset Kind Image data
Dataset Size 55.9 MB
4D view
SSBD:OMERO
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# 7569
Datast ID Fig3D_PCB_125uM
Dataset Kind Image data
Dataset Size 40.7 MB
4D view
SSBD:OMERO
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# 7570
Dataset Kind Image data
Dataset Size 50.8 MB
4D view
SSBD:OMERO
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# 7571
Dataset Kind Image data
Dataset Size 61.0 MB
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SSBD:OMERO
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# 7572
Datast ID Fig7D_WT_BV_125uM
Dataset Kind Image data
Dataset Size 50.8 MB
4D view
SSBD:OMERO
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# 7573
Datast ID Fig7D_WT_PCB_125uM
Dataset Kind Image data
Dataset Size 50.8 MB
4D view
SSBD:OMERO
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