Thy1-GCaMP2.2c founder line 8 transgenic mice express the fluorescent calcium indicator protein GCaMP2.2c in activated neurons permitting fluorescent, neuronal imaging in live animals.
Guoping Feng, Massachusetts Institute of Technology
Genetic Background | Generation |
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Allele Type |
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Transgenic (Reporter) |
This transgenic strain contains the thymus cell antigen 1 (Thy1) promoter driving expression of an eGFP-calmodulin fusion protein, GCaMP2.2c in the central nervous system. Specifically, GCaMP is expressed in the cortex, hippocampus, thalamus, cerebellum, superior colliculus, amygdala, brain stem, retina and spinal cord. An increase of green fluorescence in specific neurons can be observed as the result of elevated intracellular calcium. In these GCaMP2.2c mice, cytoplasmic fluorescence is evident in apical dendrites, dendritic spines of layer V neurons of the living cortex. GCAMP expression is lower in these mice than in B6;CBA-Tg(Thy1-GCamp3)6Gfng/J mice (Stock No. 017893). GCaMP2.2c mice allow fluorescent detection of activated neurons at the cellular level in brain slices and live animals.
This transgene contains a calcium-sensing molecule, GCaMP2.2c, under the transcriptional control of the thymus cell antigen 1 (Thy1) promoter.
The Thy1 promoter used here is a modified regulatory region of the "murine thy1.2 gene" (extending from the promoter to the intron following exon 4, excluding exon 3 and its flanking introns). The GCaMP2 consists of, from 5' to 3', a poly-histidine tag (6XHis), 13-residue peptide of myosin light chain kinase (M13), a circularly permutated eGFP interrupted at residue 145, calmodulin (CaM), and a polyA recognition sequence. Mutations resulting in increased brightness (D180Y and V93I), thermal stability (V163A, and S175G) and GFP dimerization prevention (A206K) were introduced. To further stabilize GCaMP2 and increase fluorescence, two more mutations (R2V and S118C) were introduced, resulting in the creation of GCaMP2.2c. This construct was injected into fertilized (C57BL6/J x CBA) F1)F1 oocytes and mice from founder line 8 were bred to C57BL/6 mice to establish a colony. Transgenic offspring were backcrossed to C57BL/6 mice. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Expressed Gene | GCaMP, Genetically encoded calcium indicator, |
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Site of Expression |
Allele Name | transgene insertion 8, Guoping Feng |
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Allele Type | Transgenic (Reporter) |
Allele Synonym(s) | GCaMP2.2c |
Gene Symbol and Name | Tg(Thy1-GCaMP2.2c)8Gfng, transgene insertion 8, Guoping Feng |
Gene Synonym(s) | |
Promoter | Thy1, thymus cell antigen 1, theta, mouse, laboratory |
Expressed Gene | GCaMP, Genetically encoded calcium indicator, |
Strain of Origin | (CBA x C57BL/6J)F1 |
Chromosome | UN |
Molecular Note | This transgene contains a calcium-sensing molecule, GCaMP2.2c, under the transcriptional control of the thymus cell antigen 1 (Thy1) promoter. The Thy1 promoter used here is a modified regulatory region of the "murine thy1.2 gene" (extending from the promoter to the intron following exon 4, excluding exon 3 and its flanking introns). The GCaMP2 consists of, from 5' to 3', a poly-histidine tag (6XHis), 13-residue peptide of myosin light chain kinase (M13), a circularly permutated eGFP interrupted at residue 145, calmodulin (CaM), and a polyA recognition sequence. Mutations resulting in increased brightness (D180Y and V93I), thermal stability (V163A, and S175G) and GFP dimerization prevention (A206K) were introduced. To further stabilize GCaMP2 and increase fluorescence, two more mutations (R2V and S118C) were introduced, resulting in the creation of GCaMP2.2c. Line 8 was generated. The thymus cell antigen 1 (Thy1) promoter drives expression of an eGFP-calmodulin fusion protein, GCaMP2.2c in the central nervous system. Specifically, GCaMP is expressed in the cortex, hippocampus, thalamus, cerebellum, superior colliculus, amygdala, brain stem, retina and spinal cord. An increase of green fluorescence in specific neurons can be observed as the result of elevated intracellular calcium. In these GCaMP2.2c mice, cytoplasmic fluorescence is evident in apical dendrites, dendritic spines of layer V neurons of the living cortex. GCAMP expression is lower in these mice than in B6;CBA-Tg(Thy1-GCamp3)6Gfng/J mice (Stock No. 017893). GCaMP2.2c mice allow fluorescent detection of activated neurons at the cellular level in brain slices and live animals. |
Mutations Made By | Guoping Feng, Massachusetts Institute of Technology |
When maintaining a live colony, hemizygous mice may be bred to wildtype (non-carrier) mice from the colony or to C57BL/6J inbred mice (Stock No. 000664). The donating investigator reports that homozygotes are viable.
When using the Thy1-GCaMP2.2c line 8 mouse strain in a publication, please cite the originating article(s) and include JAX stock #017892 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Hemizygous or Non carrier for Tg(Thy1-GCaMP2.2c)8Gfng |
Frozen Mouse Embryo | B6;CBA-Tg(Thy1-GCaMP2.2c)8Gfng/J | $2595.00 |
Frozen Mouse Embryo | B6;CBA-Tg(Thy1-GCaMP2.2c)8Gfng/J | $2595.00 |
Frozen Mouse Embryo | B6;CBA-Tg(Thy1-GCaMP2.2c)8Gfng/J | $3373.50 |
Frozen Mouse Embryo | B6;CBA-Tg(Thy1-GCaMP2.2c)8Gfng/J | $3373.50 |
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