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A bright and high-performance genetically encoded Ca2+ indicator based on mNeonGreen fluorescent protein

Landon Zarowny, View ORCID ProfileAbhi Aggarwal, Virginia M.S. Rutten, Ilya Kolb, The GENIE Project, Ronak Patel, Hsin-Yi Huang, Yu-Fen Chang, Tiffany Phan, Richard Kanyo, Misha Ahrens, View ORCID ProfileW. Ted Allison, Kaspar Podgorski, Robert E. Campbell
doi: https://doi.org/10.1101/2020.01.16.909291
Landon Zarowny
1Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada
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Abhi Aggarwal
1Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
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  • ORCID record for Abhi Aggarwal
Virginia M.S. Rutten
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
3Gatsby Computational Neuroscience Unit, UCL, London, UK
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Ilya Kolb
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
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2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
Ronak Patel
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
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Hsin-Yi Huang
4LumiSTAR Biotechnology, Inc. National Biotechnology Research Park, Taipei City 115, Taiwan
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Yu-Fen Chang
4LumiSTAR Biotechnology, Inc. National Biotechnology Research Park, Taipei City 115, Taiwan
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Tiffany Phan
1Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada
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Richard Kanyo
5Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada
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Misha Ahrens
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
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W. Ted Allison
5Department of Biological Sciences, University of Alberta, Edmonton, Alberta, Canada
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Kaspar Podgorski
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia, United States
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Robert E. Campbell
1Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada
6Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo, Japan
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  • For correspondence: robert.e.campbell@ualberta.ca
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Abstract

Genetically encodable calcium ion (Ca2+) indicators (GECIs) based on green fluorescent proteins (GFP) are powerful tools for imaging of cell signaling and neural activity in model organisms. Following almost two decades of steady improvements in the Aequorea victoria GFP (avGFP)-based GCaMP series of GECIs, the performance of the most recent generation (i.e., GCaMP7) may have reached its practical limit due to the inherent properties of GFP. In an effort to sustain the steady progression towards ever-improved GECIs, we undertook the development of a new GECI based on the bright monomeric GFP, mNeonGreen (mNG). The resulting indicator, mNG-GECO1, is 60% brighter than GCaMP6s in vitro and provides comparable performance as demonstrated by imaging Ca2+ dynamics in cultured cells, primary neurons, and in vivo in larval zebrafish. These results suggest that mNG-GECO1 is a promising next-generation GECI that could inherit the mantle of GCaMP and allow the steady improvement of GECIs to continue for generations to come.

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Posted January 21, 2020.
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A bright and high-performance genetically encoded Ca2+ indicator based on mNeonGreen fluorescent protein
Landon Zarowny, Abhi Aggarwal, Virginia M.S. Rutten, Ilya Kolb, The GENIE Project, Ronak Patel, Hsin-Yi Huang, Yu-Fen Chang, Tiffany Phan, Richard Kanyo, Misha Ahrens, W. Ted Allison, Kaspar Podgorski, Robert E. Campbell
bioRxiv 2020.01.16.909291; doi: https://doi.org/10.1101/2020.01.16.909291
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A bright and high-performance genetically encoded Ca2+ indicator based on mNeonGreen fluorescent protein
Landon Zarowny, Abhi Aggarwal, Virginia M.S. Rutten, Ilya Kolb, The GENIE Project, Ronak Patel, Hsin-Yi Huang, Yu-Fen Chang, Tiffany Phan, Richard Kanyo, Misha Ahrens, W. Ted Allison, Kaspar Podgorski, Robert E. Campbell
bioRxiv 2020.01.16.909291; doi: https://doi.org/10.1101/2020.01.16.909291

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