This Orai1 knock-out strain is useful in studies of regulation of cytosolic and endoplasmic reticulum calcium levels, store-operated calcium channels and T cell and B cell function.
Anjana Rao, La Jolla Institute for Allergy and Immunology
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Null/Knockout) | Orai1 | ORAI calcium release-activated calcium modulator 1 |
The targeted Orai1 gene encodes a pore subunit of the store-operated calcium channel, CRAC (calcium release-activated calcium) channel. Mutations in this gene have been associated with primary immunodeficiency-9 and tubular aggregate myopathy-2. These knock-out mice carry an allele of Orai1 in which exon 1 has been excised.
No gene product (mRNA) is detected by RT-PCR analysis of CD4+ T cells and CD19+ B cells from homozygotes. Fewer than expected homozygotes on the C57BL/6 background are born, and homozygotes are perinatal lethal, with all homozygous pups dying by 1.5 days after birth.
MEFs isolated from homozygotes exhibit significantly impaired store-operated Ca2+ influx.
A targeting vector was designed to disrupt exon 1 of the targeted gene with a loxP site flanked self-deleting selection cassette (containing a neomycin resistance gene and Cre recombinase under the control of a testes-specific angiotensin-converting enzyme promoter). The construct was electroporated into C57BL/6NTac derived B6/3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The resulting chimeric animals were bred to C57BL/6J to achieve germline transmission and allow the excision of the self-deleting selection cassette.
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).
Allele Name | targeted mutation 1, Anjana Rao |
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Allele Type | Targeted (Null/Knockout) |
Allele Synonym(s) | |
Gene Symbol and Name | Orai1, ORAI calcium release-activated calcium modulator 1 |
Gene Synonym(s) | |
Strain of Origin | C57BL/6NTac |
Chromosome | 5 |
General Note | ES cell line = B6/3. |
Molecular Note | Exon 1 was replaced with a self-deleting neomycin cassette that was excised in the germ line of male mice carrying the targeted allele. Gene inactivation was confirmed by a lack mRNA transcript in CD4+ T cells and B cells. No protein expression was detected in the skin by immunohistochemical analysis. |
When maintaining a live colony, heterozygous mice may be bred to wildtype siblings, or to C57BL/6J inbred mice (Stock No. 000664). Homozygotes are perinatal lethal.
When using the Orai1 KO mouse strain in a publication, please cite the originating article(s) and include JAX stock #023352 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
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Heterozygous or wildtype for Orai1<tm1Rao>/ |
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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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