pCAGGS-GCaMP2 transgenic mice are a fluorescent calcium indicator strain that exhibits high calcium-sensor expression in widespread cells/tissues. Upon GCaMP2 binding to calcium, increased EGFP fluorescence is observed. The entire transgene is flanked with loxP sites.
Michael I Kotlikoff, Cornell University
Genetic Background | Generation |
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Allele Type |
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Transgenic (Reporter) |
pCAGGS-GCaMP2 transgenic mice express the fluorescent calcium indicator GCaMP2 under control of the CAG hybrid promoter. pCAGGS-GCaMP2 transgenic mice from founder line 2 exhibit high sensor expression (GCaMP2 expression/EGFP fluorescence) in widespread cells/tissues.
In the absence of calcium binding, low/baseline EGFP fluorescence is observed. Following calcium binding (such as muscle contraction, arteriolar vasodilation etc.), bright EGFP fluorescence is observed. Of note, the entire transgene is flanked with loxP sites, allowing for removal of the transgene via Cre recombinase if so desired.
Hemizygous and homozygous mice are viable and fertile with no reported gross physical or behavioral abnormalities. The donating investigator reports that homozygotes have brighter fluorescence than hemizygotes.
Calcium is a key molecular signal for cell functions including heart, smooth muscle, vessel, and airway contraction, lung secretion, autonomic neurotransmission and immunocyte function. pCAGGS-GCaMP2 transgenic mice may be useful to examine calcium signaling in widespread cells/tissues both in vivo and in vitro.
The genetically encoded calcium indicator GCaMP2 is a calcium-sensing molecule composed of a chicken smooth muscle M13 fragment of myosin light chain kinase, a circularly permutated EGFP (with several amino acid substitutions designed to increase brightness, dynamic range and thermal stability, as well as to prevent GFP dimerization), and a rat calmodulin sequence. In the absence of calcium binding, dim EGFP fluorescence is expected as there is a pore from the outside of its barrel into the chromophore. Upon calcium binding, this pore becomes occupied and fluorescence is increased.
This mouse model is available by way of a collaborative effort between Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (CHROMus) and The Jackson Laboratory.
pCAGGS-GCaMP2 transgenic mice were designed in the laboratory of Dr. Michael I. Kotlikoff (Cornell University) as part of Cornell/National Heart Lung Blood Resource for Optogenetic Mouse Signaling (CHROMus).
The 3.7 kbp pCAGGS-GCaMP2 transgene has a loxP site, the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), the GCaMP2 sequence (described below), a polyadenylation signal and a second loxP site.
The transgene was microinjected into B6D2F2 embryos, and founder animals were bred to C57BL/6J inbred mice for germline transmission. pCAGGS-GCaMP2 founder line 2 was identified with high sensor expression. The donating investigator reports that pCAGGS-GCaMP2 transgenic mice from this founder line were backcrossed two generations to C57BL/6J wildtype mice, then bred together nine generations as homozygotes, and then backcrossed five more generations to C57BL/6J wildtype mice.
In 2015, mice were sent to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To establish our living colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J inbred females (Stock No. 000664).
Of note, the donating investigator reports that, at least once during backcrossing, a hemizygous female was bred to a C57BL/6J inbred male (thus the Y chromosome of the congenic strain is of C57BL/6J origin).
The genetically encoded calcium indicator GCaMP2 is a calcium-sensing molecule composed of a 35 aa polyHis plasmid leader sequence (RSET; essential for thermal stability), a 13 residue peptide of chicken smooth muscle myosin light chain kinase (M13), a circularly permutated EGFP (cpEGFP; aa 149-238 followed by aa 1-144 [with mutations listed below]), and a rat calmodulin DNA fragment (CaM; aa 2-148). The cpEGFP mutations result in increased brightness (D180Y and V93I), thermal stability (V163A and S175G) and GFP dimerization prevention (A206K).
Expressed Gene | GCaMP, Genetically encoded calcium indicator, |
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Site of Expression | EGFP expression is widespread. |
Allele Name | transgene insertion 2, Michael I Kotlikoff |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | pCAGGS-GCaMP2 |
Gene Symbol and Name | Tg(CAG-GCaMP2)2Mik, transgene insertion 2, Michael I Kotlikoff |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | GCaMP, Genetically encoded calcium indicator, |
Site of Expression | EGFP expression is widespread. |
Strain of Origin | (C57BL/6 x DBA/2)F2 |
Chromosome | UN |
Molecular Note | This 3.7 kbp transgene has a loxP site, the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (from pCAGGS), the GCaMP2 sequence (described below), a polyadenylation signal and a second loxP site. Founder line 2 was identified with high sensor expression. The genetically encoded calcium indicator GCaMP2 is a calcium-sensing molecule composed of a 35 aa polyHis plasmid leader sequence (RSET; essential for thermal stability), a 13 residue peptide of chicken smooth muscle myosin light chain kinase (M13), a circularly permutated EGFP (cpEGFP; aa 149-238 followed by aa 1-144 [with mutations listed below]), and a rat calmodulin DNA fragment (CaM; aa 2-148). The cpEGFP mutations result in increased brightness (D180Y and V93I), thermal stability (V163A and S175G) and GFP dimerization prevention (A206K). |
When maintaining a live colony, hemizygous mice may be bred to wildtype (noncarrier) mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). Alternatively, homozygous mice may be bred together.
When using the CHROMus line pCAGGS-GCaMP2 mouse strain in a publication, please cite the originating article(s) and include JAX stock #025619 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemiztgous or non carrier for Tg(CAG-GCaMP2)2Mik |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-GCaMP2)2Mik/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-GCaMP2)2Mik/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-GCaMP2)2Mik/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6.Cg-Tg(CAG-GCaMP2)2Mik/J Frozen Embryo | $3373.50 |
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