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Rational design of a high-affinity, fast, red calcium indicator R-CaMP2

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Rational design of a high-affinity, fast, red calcium indicator R-CaMP2

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  • Author: Masatoshi Inoue, Atsuya Takeuchi, Shin-ichiro Horigane, Masamichi Ohkura, Keiko Gengyo-Ando, Hajime Fujii, Satoshi Kamijo, Sayaka Takemoto-Kimura, Masanobu Kano, Junichi Nakai, Kazuo Kitamura, Haruhiko Bito
Fluorescent Ca2+ reporters are widely used as readouts of neuronal activities. Here we designed R-CaMP2, a high-affinity red genetically encoded calcium indicator (GECI) with a Hill coefficient near. Use of the calmodulin-binding sequence of CaMKK-α and CaMKK-β in lieu of an M13 sequence resulted in threefold faster rise and decay times of Ca2+ transients than R-CaMP1.07. These features allowed resolving single action potentials (APs) and recording fast AP trains up to 20–40 Hz in cortical slices. Somatic and synaptic activities of a cortical neuronal ensemble in vivo were imaged with similar efficacy as with previously reported sensitive green GECIs.

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