jRGECO1 - Improved Red Protein Indicators for In Vivo Calcium Imaging

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Reagents

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jRGECO1 - Improved Red Protein Indicators for In Vivo Calcium Imaging

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Calcium Indicators

Optical imaging of calcium dynamics using genetically encoded calcium indicators (GECIs) is a powerful tool for systems neuroscience. The current state-of-the-art GECIs emit green light (green GECIs) and are derived from Green Fluorescent Protein (GFP). Still, green fluorescence fails in several important experimental setups where tissue absorption is of concern, creating a need for red indicators.

Red GECIs may be used for:

Deep tissue imaging
Dual-color imaging
Parallel use with light-sensitive ion channel (ChR2)

Joint efforts at Janelia have generated high-sensitivity red GECIs based on RCaMP and R-GECO for in vivo imaging of neural activity. New mRuby-based RCaMP (jRCaMP GENIE Page; jRCaMP Tool Page), and mApple-based R-GECO (jRGECO1 GENIE page), available below, variants have improved sensitivity and kinetics. The sensitivity of jRGECO1a in vivo was similar to state-of-the-art GFP-based GCaMP6. The performance of both reagents has been characterized in cultured neurons and in mice, Drosophila, zebrafish, and C. elegans in vivo. These reagents are available and distributed through Addgene.org as plasmids, Penn Vector Core as viral vectors, and in flies from the Bloomington Stock Center.

Opportunity

Available for non-profit research from Addgene. Available for purchase from Vigene Biosciences. Commercial licenses are also available for internal research or for distribution.

jRGECO1 calcium indicators

Plasmids available from Addgene.org:

Name	Description	Addgene Plasmid ID	Reference
CMV-NES-jRGECO1a	red, improved SNR	61563	Dana et al., 2016

Viruses available from University of Pennsylvania Viral Vector Core:

Name	Penn Catalog Number	Reference
AAV1.Syn.NES.jRGECO1a.WPRE.SV40	AV-1-PV3849	Dana et al., 2016
AAV1.CAG.Flex.NES.jRGECO1a.WPRE.SV40	not yet available	Dana et al., 2016
AAV1.Syn.Flex.NES.jRGECO1a.WPRE.SV40	AV-1-PV3848	Dana et al., 2016
AAV1.Syn.NES.jRGECO1b.WPRE.SV40	AV-1-PV3846	Dana et al., 2016
AAV1.CAG.Flex.NES.jRGECO1b.WPRE.SV40	not yet available	Dana et al., 2016
AAV1.Syn.Flex.NES.jRGECO1b.WPRE.SV40	not yet available	Dana et al., 2016

Mice available from The Jackson Laboratory:

Name	Description	Jax Stock Number	Reference
Thy1-jRGECO1a-WPRE (founder line GP8.20)	red fluorescent calcium indicator with high sensitivity detection of action potentials, variable neuronal expression pattern	030525	Dana et al., manuscript in preparation
Thy1-jRGECO1a-WPRE (founder line GP8.31)	red fluorescent calcium indicator with high sensitivity detection of action potentials, variable neuronal expression pattern	030526	Dana et al., manuscript in preparation
Thy1-jRGECO1a-WPRE (founder line GP8.58)	red fluorescent calcium indicator with high sensitivity detection of action potentials, variable neuronal expression pattern	030527	Dana et al., manuscript in preparation
Thy1-jRGECO1a-WPRE (founder line GP8.62)	red fluorescent calcium indicator with high sensitivity detection of action potentials, variable neuronal expression pattern	030528	Dana et al., manuscript in preparation