Gpr65-ires-Cre knock-in mice express Cre recombinase under the direction of the Gpr65 promoter in vagal sensory neurons in the gastrointestinal tract that respond to ingested nutrients.
Stephen Liberles, Harvard Medical School
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing) | Gpr65 | G-protein coupled receptor 65 |
Gpr65-ires-Cre mice have an IRES-Cre fusion protein inserted 3 base pairs downstream of the stop codon of the G-protein coupled receptor 65 (Gpr65) gene. The presence of the IRES-Cre cassette downstream of the stop codon allows for expression of GPR65 and cre. GPR65 encodes a receptor present on several immune cell subsets, and also on a subset of vagal sensory neurons in the gastrointestinal tract that respond to ingested nutrients and control gut constriction and motility. The Gpr65-ires-Cre reporter is expressed in 88% of Gpr65 neurons when bred to a cre-dependent tdTomato reporter (B6;129S6-Gt(ROSA)26Sortm14(CAG-tdTomato)Hze/J Stock No. 007908) . Homozygous mice are viable and fertile. When these mice are bred with mice containing loxP-flanked sequence, Cre-mediated recombination will result in deletion of the floxed sequences in the Cre-expressing cells of the offspring.
When Gpr65-ires-Cre mice were bred to B6;129S-Gt(ROSA)26Sortm32(CAG-COP4*H134R/EYFP)Hze/J (Stock No. 012569) mice, which express a cre-inducible improved channelrhodopsin-2/EYFP fusion protein, optogenetic activation of GPR65 neurons results in stopped or reduced gastric contractions without impacting breathing or heart rate, which are also under vagal control.
A targeting construct was designed by Stephen Liberles (Harvard Medical School) to insert an internal ribosomal entry site (IRES) fused to Cre Recombinase with a polyadenylation signal, followed by a frt-flanked neomycin resistance (neo) cassette, 3 base pairs downstream of the stop codon of the G-protein coupled receptor 65 (Gpr65) gene. This construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the resulting chimeric males were bred to B6;SJL-Tg(ACTFLPe)9205Dym/J mice (Stock No. 003800) to remove the neo cassette. Offspring were crossed to remove the flpe-expressing transgene, and resulting Gpr65-ires-Cre mice were bred to C57BL/6J mice for at least 2 generations. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
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Site of Expression | Gpr65 encodes a receptor present on several immune cell subsets, and also on a subset of vagal sensory neurons in the gastrointestinal tract. |
Allele Name | trageted mutation 1.1, Stephen Liberles |
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Allele Type | Targeted (Recombinase-expressing) |
Allele Synonym(s) | Gpr65-ires-Cre |
Gene Symbol and Name | Gpr65, G-protein coupled receptor 65 |
Gene Synonym(s) | |
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
Site of Expression | Gpr65 encodes a receptor present on several immune cell subsets, and also on a subset of vagal sensory neurons in the gastrointestinal tract. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 12 |
Molecular Note | A cassette encoding IRES, Cre recombinase, and an FRT-flanked neomycin selection cassette is inserted into the mouse Gpr65 locus 3 base pairs downstream of the coding sequence stop codon. |
When maintaining a live colony homozygous mice may be bred together.
When using the Gpr65-ires-Cre mouse strain in a publication, please cite the originating article(s) and include JAX stock #029282 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous for Gpr65<tm1.1(cre)Lbrl> |
Frozen Mouse Embryo | STOCK Gpr65<tm1.1(cre)Lbrl>/RcngJ | $2595.00 |
Frozen Mouse Embryo | STOCK Gpr65<tm1.1(cre)Lbrl>/RcngJ | $2595.00 |
Frozen Mouse Embryo | STOCK Gpr65<tm1.1(cre)Lbrl>/RcngJ | $3373.50 |
Frozen Mouse Embryo | STOCK Gpr65<tm1.1(cre)Lbrl>/RcngJ | $3373.50 |
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