B6;129P2-Gt(ROSA)26Sor^{tm1(Trpv1,ECFP)Mde}/J

Stock number: 008513
Research areas: Neurobiology Research, Research Tools

Description

A loxP-flanked neomycin cassette blocks expression of the rat Trpv1 (transient receptor potential cation channel, subfamily V, member 1) gene driven by the Gt(ROSA)26Sor gene in this targeted mutation/knock-in strain. Upon crossing to a tissue-specific Cre-expressing strain, TRPV1 and enhanced cyan fluorescent protein (ECFP) is expressed from the ROSA locus. Cells expressing TRPV1 are sensitive to capsaicin and similar chemical agonists. Treatment of mice or cells that have undergone Cre excision to remove the neomycin cassette can induce strong inward currents, trigger action potentials and activate stereotyped behaviors, allowing cell-type specific chemical genetic control of neuronal activity in vitro and in vivo. These mice may also be useful in studies of TCR signalling in T cells.

Detailed description

A loxP-flanked neomycin cassette blocks expression of the rat Trpv1 (transient receptor potential cation channel, subfamily V, member 1) gene driven by the Gt(ROSA)26Sor gene in this targeted mutation/knock-in strain. Upon crossing to a tissue-specific Cre-expressing strain, TRPV1 and enhanced cyan fluorescent protein (ECFP) is expressed from the ROSA locus. Cells expressing TRPV1 are sensitive to capsaicin and similar chemical agonists. T cells express functional TRPV1, which functions in TCR signaling. Treatment of mice or cells that have undergone Cre excision to remove the neomycin cassette can induce strong inward currents, trigger action potentials and activate stereotyped behaviors, allowing cell-type specific chemical genetic control of neuronal activity in vitro and in vivo. Mice that are homozygous for the floxed allele are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities.

When bred to a strain with widespread expression of Cre recombinase (see Stock No. 002858 for example), this mutant mouse strain may be useful in studies of neuronal activity.

Development

A targeting vector was used to insert a loxP-flanked neomycin cassette, rat Trpv1 (transient receptor potential cation channel, subfamily V, member 1) cDNA, and IRES-ECFP sequence into the Gt(ROSA)26Sor gene. The mutation was created in 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were backcrossed to C57BL/6 (see SNP notes below) three times by the donating laboratory, and then males were sent to The Jackson Laboratory Repository in 2008. 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). Then, the colony was backcrossed to C57BL/6J at least one more generation. C57BL/6-congenic mice from our colony were distributed from October 2009 through December 2011. From 2012 to present (December 2015), the strain has been available by rederivation from the stocks frozen in 2008.

In 2009, a 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 1 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a C57BL/6N genetic background.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(Trpv1,ECFP)Mde}
Name: targeted mutation 1, Michael D Ehlers
MGI accession ID: MGI:3796847
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Chromosome: 6
Strain of origin: 129P2/OlaHsd
Expressed genes: CFP,Cyan Fluorescent Protein,jellyfish; Trpv1,transient receptor potential cation channel, subfamily V, member 1,rat
Site of expression: A floxed Neo prevents transcription of ECFP and rat Trpv1 (transient receptor potential cation channel, subfamily V, member 1)) until mice are bred to a Cre strain. Resulting offspring express ECFP in a pattern dictated by the Cre promoter.
Molecular note: A targeting vector was used to insert a loxP-flanked neomycin cassette, rat Trpv1 (transient receptor potential cation channel, subfamily V, member 1) cDNA, and IRES-ECFP sequence into the Gt(ROSA)26Sor locus.