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GCaMP

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GCaMP

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Next-generation genetically encoded calcium indicators

Current Iterations: GCaMP3, GCaMP5, GCaMP6, jGCaMP7

Fluorescent calcium indicators are an important tool used to detect and measure calcium, an essential regulator of cellular signaling. These small molecule or protein indicators may be used to measure the activity of G-protein coupled receptors (GPCR) and ion channels, proteins commonly targeted by pharmaceuticals. Moreover, calcium indicators can be used to image and measure changes in Ca2+ concentrations associated with neural activity.

To date, the choice of calcium indicators has required a compromise based on experimental goals. Small-molecule synthetic dyes are the most sensitive and rapid indicators, but these dyes require invasive chemical or physical delivery methods, are difficult to target to specific cell types and subcellular locations, and do not persist when used in vivo. Conversely, genetically encoded calcium indicators (GECIs) can be targeted to specific cell types and imaged in a relatively non-invasive manner over long time scales, but suffer from limited sensitivity and slower kinetics.

Janelia scientists have developed a new generation of GECIs that overcome these barriers, displaying sensitivity and kinetics similar to synthetic dyes. The GCaMP6 family of GECIs is a collection of ultrasensitive, green fluorescent indicator proteins that enable reliable detection of single action potential responses in vivo and facilitate the measurement of synaptic calcium signals. Variants with improved kinetic response will be useful for studying high frequency neuronal activity, offering the fastest observed kinetics of any protein sensor for cytoplasmic Ca2+. The jGCaMP7 family has improved on GCaMP6 performance even more, with faster kinetics and more sensitive detection. 

Advantages:

  • Sensitivity, dynamic range, and kinetics that can exceed synthetic indicators and show significant improvement over existing GECIs
  • Several GCaMP6 and jGCaMP7 variants to select from with overall brightness, rise/decay kinetics, and calcium affinity tuned for diverse applications

Applications:

  • Neuroscience research using in vitro or in vivo models
  • Long-term functional imaging of neuronal activity and correlation with animal behavior
  • Screening for G-protein coupled receptor (GPCR) or ion channel agonists and antagonists

Patent Status:

Issued U.S. Patents:  8,629,2569,518,980 and  9,488,642 and 9,945,844

Opportunity:

  • Purchase viral constructs from Vigene Biosciences.
  • Obtain the plasmid from Addgene for Non-Profit Research. 
  • Commercial Licenses are available for internal research or sale. 

For inquiries, please reference:

Janelia 2009-009 (GCaMP3), Janelia 2013-001 (GCaMP5 and GCaMP6), and Janelia 2018-002

jGCaMP7 calcium indicators

Plasmids available from Addgene.org:

Name Description Addgene Plasmid ID Reference

pCMV-jGCaMP7s

improved SNR, slow kinetics

104463

Dana et al., in preparation

pCMV-jGCaMP7f

improved SNR, fast kinetics

104483

Dana et al., in preparation

pCMV-jGCaMP7b

improved SNR, higher baseline fluorescence

104484

Dana et al., in preparation

pCMV-jGCaMP7c very low baseline fluorescence 105320 Dana et al., in preparation
pAAV-syn-jGCaMP7s-WPRE improved SNR, slow kinetics 104487 Dana et al., in preparation
pAAV-syn-jGCaMP7f-WPRE improved SNR, fast kinetics 104488 Dana et al., in preparation
pAAV-syn-jGCaMP7b-WPRE improved SNR, higher baseline fluorescence 104489 Dana et al., in preparation
pAAV-syn-jGCaMP7c-WPRE very low baseline fluorescence 105321 Dana et al., in preparation
pAAV-syn-FLEX-jGCaMP7s-WPRE improved SNR, slow kinetics, Cre-dependent expression 104491 Dana et al., in preparation
pAAV-syn-FLEX-jGCaMP7f-WPRE improved SNR, fast kinetics, Cre-dependent expression 104492 Dana et al., in preparation
pAAV-syn-FLEX-jGCaMP7b-WPRE improved SNR, higher baseline fluorescence, Cre-dependent expression 104493 Dana et al., in preparation
pAAV-syn-FLEX-jGCaMP7c-WPRE very low baseline fluorescence, Cre-dependent expression 105322 Dana et al., in preparation
pAAV-CAG-FLEX-jGCaMP7s-WPRE improved SNR, slow kinetics, Cre-dependent expression 104495 Dana et al., in preparation
pAAV-CAG-FLEX-jGCaMP7f-WPRE improved SNR, fast kinetics, Cre-dependent expression 104496 Dana et al., in preparation
pAAV-CAG-FLEX-jGCaMP7b-WPRE improved SNR, higher baseline fluorescence, Cre-dependent expression 104497 Dana et al., in preparation
pAAV-CAG-FLEX-jGCaMP7c-WPRE very low baseline fluorescence, Cre-dependent expression 105323 Dana et al., in preparation

 

Viruses available from  Addgene:

Name Addgene Catalog Number Reference

AAV1-Syn-jGCaMP7s-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-jGCaMP7f-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-jGCaMP7b-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-jGCaMP7c-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-FLEX-jGCaMP7s-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-FLEX-jGCaMP7f-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-FLEX-jGCaMP7b-WPRE-SV40

In production

Dana et al., in preparation

AAV1-Syn-FLEX-jGCaMP7c-WPRE-SV40

In production

Dana et al., in preparation

AAV1-CAG-FLEX-jGCaMP7s-WPRE-SV40

In production

Dana et al., in preparation

AAV1-CAG-FLEX-jGCaMP7f-WPRE-SV40

In production

Dana et al., in preparation

AAV1-CAG-FLEX-jGCaMP7b-WPRE-SV40 In production Dana et al., in preparation
AAV1-CAG-FLEX-jGCaMP7c-WPRE-SV40 In production Dana et al., in preparation

 

Flies available from Bloomington Stock Center:

Line name Insertion Site Bloomington Stock Number Reference Chr
P{20XUAS-IVS-Syn21-op1-jGCaMP7s-p10} su(Hw)attP5 Not yet uploaded Dana et al., in preparation 2
P{20XUAS-IVS-Syn21-op1-jGCaMP7f-p10} su(Hw)attP5 Not yet uploaded Dana et al., in preparation 2
P{20XUAS-IVS-Syn21-op1-jGCaMP7b-p10} su(Hw)attP5 Not yet uploaded Dana et al., in preparation 2
P{20XUAS-IVS-Syn21-op1-jGCaMP7c-p10} su(Hw)attP5 Not yet uploaded Dana et al., in preparation 2
P{20XUAS-IVS-Syn21-op1-jGCaMP7s-p10} VK00005 79032 Dana et al., in preparation 3
P{20XUAS-IVS-Syn21-op1-jGCaMP7f-p10} VK00005 79031 Dana et al., in preparation 3
P{20XUAS-IVS-Syn21-op1-jGCaMP7b-p10} VK00005 79029 Dana et al., in preparation 3
P{20XUAS-IVS-Syn21-op1-jGCaMP7c-p10} VK00005 79030 Dana et al., in preparation 3
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7s-p10} su(Hw)attP8 Not yet uploaded Dana et al., in preparation X
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7f-p10} su(Hw)attP8 Not yet uploaded Dana et al., in preparation X
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7b-p10} su(Hw)attP8 Not yet uploaded Dana et al., in preparation X
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7c-p10} su(Hw)attP8 Not yet uploaded Dana et al., in preparation X
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7s-p10} VK00005 Not yet deposited Dana et al., in preparation 3
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7f-p10} VK00005 Not yet deposited Dana et al., in preparation 3
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7b-p10} VK00005 Not yet deposited Dana et al., in preparation 3
P{13XLexAop2-IVS-Syn21-op1-jGCaMP7c-p10} VK00005 Not yet deposited Dana et al., in preparation 3

 

GCaMP3 calcium indicators

Plasmids available from Addgene.org:

Name Description Addgene Plasmid ID Reference

​CMV-GCaMP3

Green calcium indicator

22692

Tian et al., 2009

 

Viruses available from University of Pennsylvania Viral Vector Core:

Name Penn Catalog Number Penn Map and Sequence Number Reference
AAV1.hSynap.GCaMP3.WPRE.SV40

AV-1-PV1627

Penn Vector P1627

Tian et al., 2013

AAV5.hSynap.GCaMP3.WPRE.SV40

AV-5-PV1627

Penn Vector P1627

Tian et al., 2013

AAV9.hSynap.GCaMP3.WPRE.SV40

AV-9-PV1627

Penn Vector P1627

Tian et al., 2013

AAV1.hSynap.Flex.GCaMP3.WPRE.SV40

AV-1-PV1921

Penn Vector P1921

Tian et al., 2013

AAV5.hSynap.Flex.GCaMP3.WPRE.SV40

AV-5-PV1921

Penn Vector P1921

Tian et al., 2013

AAV9.hSynap.Flex.GCaMP3.WPRE.SV40

AV-9-PV1921

Penn Vector P1921

Tian et al., 2013

 

Mice available from The Jackson Laboratory:

Name Jax Stock Number Reference

Rosa26-CAG-lox-stop-lox-GCaMP3-WPRE

014538

Zariwala et al., 2010

 

Flies available from Bloomington Stock Center:

Name Insertion Site Bloomington Stock Number Reference Chr

P{UAS-GCaMP3.T}

attP40

32116

Tian et al., 2009

 2

P{20XUAS-GCaMP3} su(Hw)attP8 32234 Tian et al., 2009  1
P{20XUAS-GCaMP3} attP18 32235 Tian et al., 2009  1
P{20XUAS-GCaMP3} attP2 32236 Tian et al., 2009  3
PBac{20XUAS-GCaMP3} VK00005 32237 Tian et al., 2009  3

GCaMP5 calcium indicators

Plasmids available from Addgene.org:

Name Description Addgene Plasmid ID Reference

CMV-GCaMP5G

Green calcium indicator

31788

Akerboom et al., 2012

 

Viruses available from University of Pennsylvania Viral Vector Core:

Name Penn Catalog Number Penn Map and Sequence Number Reference

AAV1.hSynap.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-1-PV2478

Penn Vector P2478

Akerboom et al., 2013

AAV5.hSynap.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-5-PV2478

Penn Vector P2478

Akerboom et al., 2013

AAV9.hSynap.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-9-PV2478

Penn Vector P2478

Akerboom et al., 2013

AAV9.CAG.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-9-PV2643

Penn Vector P2643

Akerboom et al., 2013

AAV1.CAG.Flex.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-1-PV2541

Penn Vector P2541

Akerboom et al., 2013

AAV9.CAG.Flex.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-9-PV2541

Penn Vector P2541

Akerboom et al., 2013

AAV1.hSynap.Flex.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-1-PV2540

Penn Vector 2540

Akerboom et al., 2013

AAV5.hSynap.Flex.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-5-PV2540

Penn Vector P2540

Akerboom et al., 2013

AAV9.hSynap.Flex.GCaMP5G(GCaMP3-T302L.R303P.D380Y).WPRE.SV40

AV-9-PV2540

Penn Vector P2540

Akerboom et al., 2013

 

Flies available from Bloomington Stock Center:

Name Insertion Site Bloomington Stock Number Reference Chr
P{20XUAS-IVS-GCaMP5G} attP40 42037 Akerboom et al., 2012  2
P{20XUAS-IVS-GCaMP5G} VK00005 42038 Akerboom et al., 2012  3

GCaMP6 calcium indicators

Plasmids available from Addgene.org:

Name Description Addgene Plasmid ID Reference

CMV-GCaMP6s

improved SNR, slower kinetics

40753

Chen et al., 2013

CMV-GCaMP6m

improved SNR, intermediate kinetics

40754

Chen et al., 2013

CMV-GCaMP6f

improved SNR, faster kinetics

40755

Chen et al., 2013

 

Viruses available from University of Pennsylvania Viral Vector Core:

Name Penn Catalog Number Penn Map and Sequence Number Reference

AAV1.Syn.GCaMP6f.WPRE.SV40

AV-1-PV2822

Penn Vector P2822

Chen et al., 2013

AAV1.Syn.GCaMP6s.WPRE.SV40

AV-1-PV2824

Penn Vector P2824

Chen et al., 2013

AAV1.Syn.GCaMP6m.WPRE.SV40

AV-1-PV2823

Penn Vector P2823

Chen et al., 2013

AAV5.Syn.GCaMP6m.WPRE.SV40

AV-5-PV2823

Penn Vector P2823

Chen et al., 2013

AAV5.Syn.GCaMP6f.WPRE.SV40

AV-5-PV2822

Penn Vector P2822

Chen et al., 2013

AAV5.Syn.GCaMP6s.WPRE.SV40

AV-5-PV2824

Penn Vector P2824

Chen et al., 2013

AAV9.Syn.GCaMP6f.WPRE.SV40

AV-9-PV2822

Penn Vector P2822

Chen et al., 2013

AAV9.Syn.GCaMP6m.WPRE.SV40

AV-9-PV2823

Penn Vector P2823

Chen et al., 2013

AAV9.Syn.GCaMP6s.WPRE.SV40

AV-9-PV2824

Penn Vector P2824

Chen et al., 2013

AAV1.CAG.GCaMP6s.WPRE.SV40

AV-1-PV2833

Penn Vector P2833

Chen et al., 2013

AAV5.CAG.GCaMP6s.WPRE.SV40

AV-5-PV2833

Penn Vector P2833

Chen et al., 2013

AAV9.CAG.GCaMP6s.WPRE.SV40

AV-9-PV2833

Penn Vector P2833

Chen et al., 2013

AAV1.CAG.Flex.GCaMP6f.WPRE.SV40

AV-1-PV2816

Penn Vector 2816

Chen et al., 2013

AAV1.CAG.Flex.GCaMP6m.WPRE.SV40

AV-1-PV2817

Penn Vector 2817

Chen et al., 2013

AAV1.CAG.Flex.GCaMP6s.WPRE.SV40

AV-1-PV2818

Penn Vector 2818

Chen et al., 2013

AAV1.Syn.Flex.GCaMP6f.WPRE.SV40

AV-1-PV2819

Penn Vector 2819

Chen et al., 2013

AAV1.Syn.Flex.GCaMP6m.WPRE.SV40

AV-1-PV2820

Penn Vector 2820

Chen et al., 2013

AAV1.Syn.Flex.GCaMP6s.WPRE.SV40

AV-1-PV2821

Penn Vector 2821

Chen et al., 2013

AAV5.CAG.Flex.GCaMP6f.WPRE.SV40

AV-5-PV2816

Penn Vector 2816

Chen et al., 2013

AAV5.CAG.Flex.GCaMP6m.WPRE.SV40

AV-5-PV2817

Penn Vector 2817

Chen et al., 2013

AAV5.CAG.Flex.GCaMP6s.WPRE.SV40

AV-5-PV2818

Penn Vector 2818

Chen et al., 2013

AAV5.Syn.Flex.GCaMP6f.WPRE.SV40

AV-5-PV2819

Penn Vector 2819

Chen et al., 2013

AAV5.Syn.Flex.GCaMP6m.WPRE.SV40

AV-5-PV2820

Penn Vector 2820

Chen et al., 2013

AAV5.Syn.Flex.GCaMP6s.WPRE.SV40

AV-5-PV2821

Penn Vector 2821

Chen et al., 2013

AAV9.CAG.Flex.GCaMP6f.WPRE.SV40

AV-9-PV2816

Penn Vector 2816

Chen et al., 2013

AAV9.CAG.Flex.GCaMP6m.WPRE.SV40

AV-9-PV2817

Penn Vector 2817

Chen et al., 2013

AAV9.CAG.Flex.GCaMP6s.WPRE.SV40

AV-9-PV2818

Penn Vector 2818

Chen et al., 2013

AAV9.Syn.Flex.GCaMP6f.WPRE.SV40

AV-9-PV2819

Penn Vector 2819

Chen et al., 2013

AAV9.Syn.Flex.GCaMP6m.WPRE.SV40

AV-9-PV2820

Penn Vector 2820

Chen et al., 2013

AAV9.Syn.Flex.GCaMP6s.WPRE.SV40

AV-9-PV2821

Penn Vector 2821

Chen et al., 2013

 

 

Mice available from The Jackson Laboratory:

Name Description Jax Stock Number Reference

Thy1-GCaMP6s-WPRE (founder line GP4.3)

improved SNR, slower kinetics, variable neuronal expression pattern

024275

Dana et al., 2015

Thy1-GCaMP6s-WPRE (founder line GP4.6) improved SNR, slower kinetics, variable neuronal expression pattern 028278 Dana et al., 2015

Thy1-GCaMP6s-WPRE (founder line GP4.12)

improved SNR, slower kinetics, variable neuronal expression pattern

025776

Dana et al., 2015

Thy1-GCaMP6f-WPRE (founder line GP5.1) improved SNR, faster kinetics, variable neuronal expression pattern 028279 Dana et al., 2015
Thy1-GCaMP6f-WPRE (founder line GP5.3) improved SNR, faster kinetics, variable neuronal expression pattern 028280 Dana et al., 2015

Thy1-GCaMP6f-WPRE (founder line GP5.5)

improved SNR, faster kinetics, variable neuronal expression pattern

024276

Dana et al., 2015

Thy1-GCaMP6f-WPRE (founder line GP5.11)

improved SNR, faster kinetics, variable neuronal expression pattern

024339

Dana et al., 2015

Thy1-GCaMP6f-WPRE (founder line GP5.12) improved SNR, faster kinetics, variable neuronal expression pattern 028281 Dana et al., 2015

Thy1-GCaMP6f-WPRE (founder line GP5.17)

improved SNR, faster kinetics, variable neuronal expression pattern

025393

Dana et al., 2015

 

 

Flies available from Bloomington Stock Center:

Line name Insertion Site Bloomington Stock Number Reference Chr
P{20XUAS-IVS-GCaMP6s} attP40 42476 Chen et al., 2013 2
P{20XUAS-IVS-GCaMP6m} attP40 42748 Chen et al., 2013 2
P{20XUAS-IVS-GCaMP6f} attP40 42747 Chen et al., 2013 2
PBac{20XUAS-IVS-GCaMP6s} VK00005 42749 Chen et al., 2013 3
PBac{20XUAS-IVS-GCaMP6m} VK00005 42750 Chen et al., 2013 3
P{13XLexAop2-IVS-GCaMP6s-SV40} su(Hw)attP1 44273 Chen et al., 2013 3
P{13XLexAop2-IVS-GCaMP6m-SV40} su(Hw)attP1 44588 Chen et al., 2013 3
P{13XLexAop2-IVS-GCaMP6s-SV40} su(Hw)attP5 44589 Chen et al., 2013 2
P{13XLexAop2-IVS-GCaMP6s-p10} su(Hw)attP1 44274 Chen et al., 2013 3
P{13XLexAop2-IVS-GCaMP6m-p10} su(Hw)attP1 44275 Chen et al., 2013 3
P{13XLexAop2-IVS-GCaMP6s-p10} su(Hw)attP5 44590 Chen et al., 2013 2
P{13XLexAop2-IVS-GCaMP6f-p10} su(Hw)attP5 44277 Chen et al., 2013 2
P{13XLexAop2-IVS-GCaMP6m-p10} VK00005 44276 Chen et al., 2013 3

 

C. elegans distributed upon request (contact Program Scientist):

Name

mec-4::nls-RSET-GCaMP6s:SL2:nls-TagRFP::unc-54utr

 

 

 

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Michael Perham
Director, Innovations and External Relations
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