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<p>Genetically encoded calcium ion (Ca<sup>2+</sup>) indicators (GECIs) are widely-used molecular tools for functional imaging of Ca<sup>2+</sup> 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 Ca<sup>2+</sup> in vitro (Δ<i>F</i>/<i>F</i><sub>0</sub> = 6 for FR-GECO1a, 18 for FR-GECO1c), are bright under both one-photon and two-photon illumination, and have high affinities (apparent <i>K</i><sub>d</sub> = 29 nM for FR-GECO1a, 83 nM for FR-GECO1c) for Ca<sup>2+</sup>. 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.</p>

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Far-red fluorescent genetically encoded calcium ion indicators

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年4月7日

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  • 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,
  • Getahun Tsegaye,
  • Arthur Tsang,
  • Glenn C. Turner,
  • Yufeng Zhao,
  • Jiahui Wu,
  • Kaspar Podgorski,
  • Alison G. Tebo,
  • Eric R. Schreiter,
  • Thomas E. Hughes,
  • Takuya Terai,
  • Marie-Eve Paquet,
  • Sean G. Megason,
  • Adam E. Cohen,
  • Yi Shen,
  • Robert E. Campbell

摘要

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.