ChAT-IRES-Cre knock-in mice express Cre recombinase in cholinergic neurons, without disrupting endogenous Chat expression. These mice may be useful in neurobiological research of motor function, learning and memory, Alzheimer's disease, and Down syndrome, as well as obesity and diabetes research.
The Jackson Laboratory Repository has two ChAT-IRES-Cre knock-in alleles from Dr. Bradford Lowell: ChAT-IRES-Cre::SV40pA::frt-neo-frt (Chattm2(cre)Lowl; Stock Nos. 006410/018957/028861) and ChAT-IRES-Cre::SV40pA::Δneo (Chattm1(cre)Lowl ; Stock No. 031661). Each allele also contains a partial neo fragment between the Cre and SV40 polyA. The donating investigator reports that the presence of the frt-flanked neo cassette may result in ectopic Cre expression, and they have never observed any ectopic expression in their ChAT-IRES-Cre::SV40pA::Δneo mice (January 2018).
Bradford B. Lowell, Beth Israel Deaconess Med Cntr (Harvard)
Genetic Background | Generation |
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N?+F31
|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Recombinase-expressing) | Chat | choline acetyltransferase |
The Jackson Laboratory Repository has two ChAT-IRES-Cre knock-in alleles from Dr. Bradford Lowell: ChAT-IRES-Cre::SV40pA::frt-neo-frt (Chattm2(cre)Lowl; Stock Nos. 006410/018957/028861) and ChAT-IRES-Cre::SV40pA::Δneo (Chattm1(cre)Lowl ; Stock No. 031661). Each allele also contains a partial neo fragment between the Cre and SV40 polyA (see below). The donating investigator reports that the presence of the frt-flanked neo cassette may result in ectopic Cre expression, and they have never observed any ectopic expression in their ChAT-IRES-Cre::SV40pA::Δneo mice (January 2018).
Mice that are homozygous for the ChAT-IRES-Cre::SV40pA::frt-neo-frt knock-in allele are viable and fertile. An "IRES-Cre" sequence is inserted downstream of the stop codon such that cre expression is controlled by the endogenous Chat gene promoter. Chat gene expression, however, is unaffected. Cre recombinase activity is reported in all cholinergic neurons. These mice may be useful for "Cre-lox" technology applications in neurobiology, including studies of motor function, learning and memory, Alzheimer's disease, and Down syndrome, and in obesity and diabetes research.
Characterization performed at The Jackson Laboratory (2019-2020) determined there is an incomplete neo sequence (224 bp) located 310 bp downstream of Cre STOP codon and 70 bp upstream of SV40 polyA sequence. While it is not specifically determined if the 224 bp neo fragment has any effect on transcript levels of Cre or endogenous Chat, the Cre ORF and Cre STOP codon are intact. The 224 bp partial neo fragment includes sequences complimentary to the forward, reverse and probe primers The Jackson Laboratory has used for its generic neo genotyping assay.
View Cre expression characterization for ChAT-IRES-Cre::SV40pA::frt-neo-frt allele (Chattm2(cre)Lowl ; Stock No. 006410).
Stock No. 006410 has the ChAT-IRES-Cre::SV40pA::frt-neo-frt allele (Chattm2(cre)Lowl) on a mixed C57BL/6;129 genetic background - its generation is described below.
A targeting vector was designed to insert an optimized internal ribosome entry site-linked Cre recombinase gene (followed by an SV40 polyA and frt-flanked neomycin resistance cassette) downstream of the stop codon of the choline acetyltransferase gene (Chat) on chromosome 14. The construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. Chimeric mice were bred to C57BL/6 mice. The resulting mutant offspring (still retaining the frt-flanked neo) were bred to wildtype siblings, and maintained as such prior to sending heterozygous males to The Jackson Laboratory Repository in 2006. Upon arrival, mice were bred once to C57BL/6 females to rederive our living colony. Thereafter, mutant mice were bred together to make this line homozygous.
Characterization performed at The Jackson Laboratory (2019-2020) determined there is an incomplete neo sequence (224 bp) located 310 bp downstream of Cre STOP codon and 70 bp upstream of SV40 polyA sequence. While it is not specifically determined if the 224 bp neo fragment has any effect on transcript levels of Cre or endogenous Chat, the Cre ORF and Cre STOP codon are intact. The 224 bp partial neo fragment includes sequences complimentary to the forward, reverse and probe primers The Jackson Laboratory has used for its generic neo genotyping assay.
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
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Site of Expression | Cre recombinase activity is reported in all cholinergic neurons. |
Allele Name | targeted mutation 2, Bradford B Lowell |
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Allele Type | Targeted (Recombinase-expressing) |
Allele Synonym(s) | ChAT-IRES-Cre |
Gene Symbol and Name | Chat, choline acetyltransferase |
Gene Synonym(s) | |
Expressed Gene | cre, cre recombinase, bacteriophage P1 |
Site of Expression | Cre recombinase activity is reported in all cholinergic neurons. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 14 |
Molecular Note | An optimized internal ribosome entry sequence (IRES) fused to cre recombinase followed by a neomycin resistance cassette flanked FRT sites was inserted after the ChAT stop codon in BAC (bMQ185k21) using homologous recombination in bacteria. A genomic ChAT fragment with 4kb homology arms on either side of the IRES-Cre insertion site was lifted out from the BAC using reverse recombineering techniques and transformed into ES cells. This allele was used to generate Chattm1(cre)Lowl. |
Mutations Made By | Bradford Lowell, Beth Israel Deaconess Med Cntr (Harvard) |
When maintaining a live colony at The Jackson Laboratory Repository, homozygous mice may be bred together.
When using the ChAT-IRES-Cre , ChAT-IRES-Cre::frt-neo-frt mouse strain in a publication, please cite the originating article(s) and include JAX stock #006410 in your Materials and Methods section.
Service/Product | Description | Price |
---|---|---|
Heterozygous for Chat<tm1(cre)Lowl> |
Frozen Mouse Embryo | B6;129S6-Chat<tm2(cre)Lowl>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129S6-Chat<tm2(cre)Lowl>/J Frozen Embryo | $2595.00 |
Frozen Mouse Embryo | B6;129S6-Chat<tm2(cre)Lowl>/J Frozen Embryo | $3373.50 |
Frozen Mouse Embryo | B6;129S6-Chat<tm2(cre)Lowl>/J Frozen Embryo | $3373.50 |
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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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