bioRxivBiophysics
A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein
Y. Shen, H. Dana, A. S. Abdelfattah, R. Patel, J. Shea, R. S. Molina, B. Rawal, V. Rancic, Y.-F. Chang, L. Wu, Y. Chen, Y. Qian, M. D. Wiens, N. Hambleton, K. Ballanyi, T. E. Hughes, M. Drobizhev, D. S. Kim, M. Koyama, E. R. Schreiter, R. E. Campbell
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.
- organism: human
- organism: mouse
- organism: zebrafish
- assay: fluorescence / live-cell microscopy
- research focus: cell type annotation
- research focus: imputation
- approach: experimental assay
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