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Far-red fluorescent genetically encoded calcium ion indicators

Author

Listed:
  • Rochelin Dalangin

    (University of Alberta
    Centre de recherche CERVO)

  • Bill Z. Jia

    (Harvard University
    Harvard Medical School)

  • Yitong Qi

    (Harvard University)

  • Abhi Aggarwal

    (Howard Hughes Medical Institute
    and Hotchkiss Brain Institute)

  • Kenryo Sakoi

    (The University of Tokyo)

  • Mikhail Drobizhev

    (Montana State University
    Montana State University)

  • Rosana S. Molina

    (Montana State University)

  • Ronak Patel

    (Howard Hughes Medical Institute)

  • Ahmed S. Abdelfattah

    (Howard Hughes Medical Institute
    Brown University)

  • Jihong Zheng

    (Howard Hughes Medical Institute)

  • Daniel Reep

    (Howard Hughes Medical Institute)

  • Jeremy P. Hasseman

    (Howard Hughes Medical Institute)

  • Yufeng Zhao

    (University of Alberta
    University of Toronto)

  • Jiahui Wu

    (University of Alberta
    University of Massachusetts)

  • Kaspar Podgorski

    (Howard Hughes Medical Institute
    Allen Institute for Neural Dynamics)

  • Alison G. Tebo

    (Howard Hughes Medical Institute)

  • Eric R. Schreiter

    (Howard Hughes Medical Institute)

  • Thomas E. Hughes

    (Montana State University
    Montana Molecular)

  • Takuya Terai

    (The University of Tokyo)

  • Marie-Eve Paquet

    (Centre de recherche CERVO
    Université Laval
    Université Laval)

  • Sean G. Megason

    (Harvard Medical School)

  • Adam E. Cohen

    (Harvard University)

  • Yi Shen

    (University of Alberta)

  • Robert E. Campbell

    (University of Alberta
    Centre de recherche CERVO
    The University of Tokyo
    Université Laval)

Abstract

Genetically encoded calcium ion (Ca2+) indicators (GECIs) are widely-used molecular tools for functional imaging of Ca2+ dynamics and neuronal activities with single-cell resolution. Here we report the design and development of two far-red fluorescent GECIs, FR-GECO1a and FR-GECO1c, based on the monomeric far-red fluorescent proteins mKelly1 and mKelly2. FR-GECOs have excitation and emission maxima at ~596 nm and ~644 nm, respectively, display large responses to Ca2+ in vitro (ΔF/F0 = 6 for FR-GECO1a, 18 for FR-GECO1c), are bright under both one-photon and two-photon illumination, and have high affinities (apparent Kd = 29 nM for FR-GECO1a, 83 nM for FR-GECO1c) for Ca2+. FR-GECOs offer sensitive and fast detection of single action potentials in neurons, and enable in vivo all-optical manipulation and measurement of cellular activities in combination with optogenetic actuators.

Suggested Citation

  • Rochelin Dalangin & Bill Z. Jia & Yitong Qi & Abhi Aggarwal & Kenryo Sakoi & Mikhail Drobizhev & Rosana S. Molina & Ronak Patel & Ahmed S. Abdelfattah & Jihong Zheng & Daniel Reep & Jeremy P. Hasseman, 2025. "Far-red fluorescent genetically encoded calcium ion indicators," Nature Communications, Nature, vol. 16(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-58485-z
    DOI: 10.1038/s41467-025-58485-z
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