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These double mutant mice express the human Fc fragment of IgE, high affinity I, receptor for; alpha polypeptide, (FCER1A), under the control of the human FCER1A promoter and carry the Fcer1atm1Knt targeted mutation. Mice that are hemizygous for the transgene and homozygous for the targeted mutation respond to experimental induction of anaphylaxis and are more sensitive to 2,4,6-tri-nitrobenzenesulfonic acid (TNBS)-induced colitis. These mutant mice exhibit increased levels of IL4 in the colon, enhanced intestinal permeability and modified fecal bacterial flora composition. This mutant mouse strain may be useful in studies of allergic response, anaphylaxis, hypersensitive immune response, and inflammatory bowel disease.
Jean-Pierre Kinet, Beth Israel Deaconess Medical Center
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Fcer1a | Fc receptor, IgE, high affinity I, alpha polypeptide |
Allele Type |
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Transgenic (Inserted expressed sequence, Humanized sequence) |
These double mutant mice express the human Fc fragment of IgE, high affinity I, receptor for; alpha polypeptide, (FCER1A), under the control of the human FCER1A promoter and carry the Fcer1atm1Knt targeted mutation. Mice that are hemizygous for the transgene and homozygous for the targeted mutation express a functioning chimeric receptor complex in which the human alpha-chain associates with the mouse beta- and gamma- chains. Transgene expression is detected on bone marrow derived cultured mast cells (BMMC), mast cells, basophils, monocytes, eosinophils, and epidermal Langerhans cells by FACS, hIgE binding and Western blot analysis and mimics both the human FCER1A expression and cell specificity pattern. The humanized receptor is estimated to be expressed approximately 3 to 5 fold higher than the endogenous mouse receptor. The double mutant mice respond to experimental induction of anaphylaxis and are more sensitive to 2,4,6-tri-nitrobenzenesulfonic acid (TNBS)-induced colitis. These mutant mice exhibit increased levels of IL4 in the colon, enhanced intestinal permeability and modified fecal bacterial flora composition. This mutant mouse strain may be useful in studies of allergic response, anaphylaxis, hypersensitive immune response, and inflammatory bowel disease.
These double mutant mice were generated by crossing transgenic mice carrying the Tg(FCER1A)1Bhk transgene with Fcer1atm1Knt targeted mutant mice. The targeted mutant allele was created using a targeting vector containing neomycin resistance and herpes simplex virus thymidine kinase genes to disrupt exon 4, which encodes an immunoglobulin domain. The construct was electroporated into 129S2/SvPas derived D3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts. The resulting chimeric animals were crossed to BALB/cJ mice, and then backcrossed to the same for 20 generations.
The transgenic strain was generated using a transgenic construct containing the entire human Fc fragment of IgE, high affinity I, receptor for; alpha polypeptide, (FCER1A), including 2.9kb of upstream sequence. The transgene was coinjected with a plasmid containing the hprt minigene. These constructs were transfected into ES65-154 (E14TG2a-165) embryonic stem (ES). ES cells containing the construct were injected into recipient C57BL/6 blastocysts to generate chimeric mice. The resulting male chimeric animals were bred to female mice carrying the Fcer1atm1Knt targeted mutation. The donating investigator reported that double mutant mice were then backcrossed to C57BL/6 (see SNP note below) for more than 8 generations before arriving at The Jackson Laboratory.
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 mixed C57BL/6J ; C57BL/6N genetic background.
Expressed Gene | FCER1A, Fc fragment of IgE receptor Ia, human |
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Site of Expression |
Allele Name | targeted mutation 1, Jean-Pierre Kinet |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | Fc epsilon RI alpha chain-; FcepsilonR1alpha-; FcepsilonRI-; Fcer1a-; Fcer1atm1Rav; FcerIa- |
Gene Symbol and Name | Fcer1a, Fc receptor, IgE, high affinity I, alpha polypeptide |
Gene Synonym(s) | |
Strain of Origin | 129 |
Chromosome | 1 |
General Note | ES cell line = D3 (129S2/SvPas) or E14TG2a (129P2/OlaHsd). |
Molecular Note | Exon 4, encoding an immunoglobulin domain, was disrupted by the insertion of a neomycin selection cassette. The absence of a functional encoded protein on the cell surface was determined by fluorescence activated cell sorting analysis of bone marrow mast cells obtained from homozygous mutant mice. |
Mutations Made By | Devin Turner, Beth Israel Deaconess Medical Center |
Allele Name | transgene insertion 1, Beverly H Koller |
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Allele Type | Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Synonym(s) | hFcepsilonR1alpha- hFcepsilonR1alpha+; hFcepsilonR1alphaTg |
Gene Symbol and Name | Tg(FCER1A)1Bhk, transgene insertion 1, Beverly H Koller |
Gene Synonym(s) | |
Promoter | FCER1A, Fc fragment of IgE receptor Ia, human |
Expressed Gene | FCER1A, Fc fragment of IgE receptor Ia, human |
Strain of Origin | 129P2/OlaHsd |
Chromosome | UN |
General Note | ES cells = ES65-154 (E14TG2a-165) cells |
Molecular Note | The transgenic construct contains the entire human Fc fragment of IgE, high affinity I, receptor for; alpha polypeptide, (FCER1A), including 2.9kb of upstream sequence. The transgene was coinjected with a plasmid containing the hprt minigene. These constructs were transfected into ES65-154 (E14TG2a-165) embryonic stem (ES). |
When maintaining a live colony, these double mutant mice may be bred as hemizygous for the transgene and homozygous for the targeted mutation.
When using the B6.Cg-Fcer1atm1Knt Tg(FCER1A)1Bhk/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #010506 in your Materials and Methods section.
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