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A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein

View ORCID ProfileYi Shen, Hod Dana, Ahmed S. Abdelfattah, Ronak Patel, Jamien Shea, Rosana S. Molina, Bijal Rawal, Vladimir Rancic, Yu-Fen Chang, Lanshi Wu, Yingche Chen, Yong Qian, View ORCID ProfileMatthew D. Wiens, Nathan Hambleton, Klaus Ballanyi, Thomas E. Hughes, Mikhail Drobizhev, View ORCID ProfileDouglas S. Kim, Minoru Koyama, Eric R. Schreiter, View ORCID ProfileRobert E. Campbell
doi: https://doi.org/10.1101/213082
Yi Shen
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Hod Dana
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
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Ahmed S. Abdelfattah
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Ronak Patel
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
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Jamien Shea
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
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Rosana S. Molina
3Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana 59717, USA.
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Bijal Rawal
4Department of Physiology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
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Vladimir Rancic
4Department of Physiology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
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Yu-Fen Chang
5LumiSTAR Biotechnology Incorporation, Nangang Dist., Taipei City 115, Taiwan.
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Lanshi Wu
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Yingche Chen
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Yong Qian
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Matthew D. Wiens
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Nathan Hambleton
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Klaus Ballanyi
4Department of Physiology, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
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Thomas E. Hughes
3Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana 59717, USA.
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Mikhail Drobizhev
3Department of Cell Biology and Neuroscience, Montana State University, Bozeman, Montana 59717, USA.
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Douglas S. Kim
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
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Minoru Koyama
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
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Eric R. Schreiter
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, Virginia 20147, USA.
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Robert E. Campbell
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
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Abstract

Genetically-encoded calcium ion (Ca2+) indicators (GECIs) are indispensable tools for measuring Ca2+ dynamics and neuronal activities in vitro and in vivo. Red fluorescent protein (RFP)-based GECIs enable multicolor visualization with blue or cyan-excitable fluorophores and combined use with blue or cyan-excitable optogenetic actuators. Here we report the development, structure, and validation of a new red fluorescent Ca2+ indicator, K-GECO1, based on a circularly permutated RFP derived from the sea anemone Entacmaea quadricolor. We characterized the performance of K-GECO1 in cultured HeLa cells, dissociated neurons, stem cell derived cardiomyocytes, organotypic brain slices, zebrafish spinal cord in vivo, and mouse brain in vivo.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted November 02, 2017.
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A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein
Yi Shen, Hod Dana, Ahmed S. Abdelfattah, Ronak Patel, Jamien Shea, Rosana S. Molina, Bijal Rawal, Vladimir Rancic, Yu-Fen Chang, Lanshi Wu, Yingche Chen, Yong Qian, Matthew D. Wiens, Nathan Hambleton, Klaus Ballanyi, Thomas E. Hughes, Mikhail Drobizhev, Douglas S. Kim, Minoru Koyama, Eric R. Schreiter, Robert E. Campbell
bioRxiv 213082; doi: https://doi.org/10.1101/213082
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A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein
Yi Shen, Hod Dana, Ahmed S. Abdelfattah, Ronak Patel, Jamien Shea, Rosana S. Molina, Bijal Rawal, Vladimir Rancic, Yu-Fen Chang, Lanshi Wu, Yingche Chen, Yong Qian, Matthew D. Wiens, Nathan Hambleton, Klaus Ballanyi, Thomas E. Hughes, Mikhail Drobizhev, Douglas S. Kim, Minoru Koyama, Eric R. Schreiter, Robert E. Campbell
bioRxiv 213082; doi: https://doi.org/10.1101/213082

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