A genetically encoded Ca2+ indicator based on circularly permutated sea anemone red fluorescent protein
Ahmed S. Abdelfattah,
Rosana S. Molina,
Matthew D. Wiens,
Thomas E. Hughes,
Douglas S. Kim,
Eric R. Schreiter,
Robert E. Campbell
Posted 02 Nov 2017
bioRxiv DOI: 10.1101/213082 (published DOI: 10.1186/s12915-018-0480-0)
Posted 02 Nov 2017
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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