Strain Name:
B6.129S6(Cg)-Casz1tm1Zel Tg(CD4-cre)1Cwi/Mmucd
Stock Number:
044013-UCD
Citation ID:
RRID:MMRRC_044013-UCD
Other Names:
CD4creCaszfl/fl

Strain Information

Casz1tm1Zel
Name: castor zinc finger 1; targeted mutation 1, Peggy Zelenka
Synonyms: Casz1fl
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 4
Alteration at locus: Knock-In
Casz1
Name: castor zinc finger 1
Synonyms: D4Ertd432e, 2410019P08Rik, Cst, castor
Type: Gene
Species: Mouse
Chromosome: 4
Alteration at locus: Knock-In
NCBI: 69743
Homologene: 9824
cre
Name: cre recombinase
Type: Gene
Species: Enterobacteria phage P1 (bacteriophage P1)
Chromosome:
Alteration at locus: Knock-In
Cd4
Name: CD4 antigen
Synonyms: Ly-4, L3T4
Type: Gene
Species: Mouse
Chromosome: 6
Alteration at locus: Knock-In
NCBI: 12504
HGNC: HGNC:1678
Homologene: 513
Tg(Cd4-cre)1Cwi
Name: transgene insertion 1, Christopher B Wilson
Synonyms: CD4CRE, CD4Cre, CD4-creTg, CD4p-Cre, CD4-Cre+, CD4-Cre, CD4cre, Tg(CD4-cre)1Cw1
Type: Transgene
Species: Multi-species
Chromosome: unknown
Alteration at locus: Transgenic (cre)
Genetic Alterations

A bacterial artificial chromosome clone of 129/Sv origin spanning the entire locus was obtained from Source BioScience Life Sciences, UK. A LoxP site 159 bp upstream of exon 9 and another 135 bp downstream of exon 11, respectively, were inserted so that a genomic region of 2.738 kb containing exons 9 through 11 are flanked by the two engineered LoxP sites. Removal of this genomic region upon Cre excision will lead to the deletion of 395 amino acid residues from the total of 1762 amino acids, which accounts for the longest transcript of the gene (NP_001152816.1)

Genotype Determination
ES Cell Line
W4 derived from 129S6/SvEvTac
Phenotype
Immune abnormalities: Casz1 deficiency in CD4+ T cells lowers susceptibility to experimental autoimmune encephalomyelitis, consistent with the reduced frequency of Th17 cells, despite an increase in Th1 cells in mice. Loss of Casz1 in the context of mucosal Candida infection severely impairs Th17 and Treg responses and lowers the ability of the mice to clear the secondary infection. Importantly, in both the models, the absence of Casz1 causes a significant diminution in IFN-γ+IL-17A+ double-positive inflammatory Th17 cells (Th1* cells) in tissues in vivo. Transcriptome analyses of CD4+ T cells lacking Casz1 show a signature consistent with defective Th17 differentiation. With regards to Th17 differentiation, Casz1 limits repressive histone marks and enables acquisition of permissive histone marks at Rorc, Il17a, Ahr, and Runx1 loci. Taken together, these data identify Casz1 as a new Th plasticity regulator having important clinical implications for autoimmune inflammation and mucosal immunity.
Strain Development
Mouse Casz1 (chr4:148,804,392-148,954,892) conditional targeting: BAC clone was genetically engineered as described above. The associated deletion will affect isoforms 1 and 2 and it is expected to leave downstream coding sequences out-of-frame. To place the two LoxP sites precisely at the designated locations, the approach of ET-recombination (or Recombineering) was used. Briefly, the upstream LoxP site was introduced by insertion of a pre-constructed Neomycin (Neo)/Kanamycin (Kan) cassette flanked by LoxP sites (driven by the E. coli promoter gb2) through ET-mediated homologous recombination in the E. coli host of the BAC clone. Correctly recombined clones were enriched by Kan selection, identified by PCR reactions detecting recombination junctions, and subjected to next round of manipulation in which the inserted LoxP-Neo-Loxp cassette was removed by Cre excision leaving one LoxP site precisely the pre-designated location. This was achieved through transforming the E. coli host with a Cre- encoding plasmid in which the Cre expression is rendered under the control of the temperature sensitive E. coli promoter cI578 (λPR promoter, GeneBridges, Heidelberg, Germany). Switching from 30C to 37C will activate the expression of Cre recombinase. After an overnight incubation at 37C, clones were subjected to PCR screening identifying correctly recombined clones and subsequent verification by sequencing for the presence of the LoxP site at the desired location. In a similar fashion, the downstream LoxP site was inserted except that Neo)/Kan cassette used in this round of manipulation is also driven by PGK (Mouse phosphoglycerate kinase 1) promoter and flanked with LoxP–Flippase recognition target (FRT) upstream and FRT only downstream. The LoxP-FRT-Neo-FRT cassette is not removed as in the case of the first Neo cassette insertion so that the neo gene driven by the PGK promoter can be used as a positive selection marker in ES. Finally, the modified BAC clone containing appropriately positioned LoxP sites and the Neo selection marker flanked by FRT sites was truncated to a more manageable size so that both upstream and downstream targeting arms were reduced to about 5 kb of homologous sequences to the target locus. This last step of manipulation was achieved through another round of ET recombination between the modified BAC and a recipient plasmid vector as described. W4 ES cell line derived from the 129S6/SvEvTac from Taconic was used.

The Casz1-floexed strain was bred with mice transgenic for CD4-cre at Case Western Reserve University to study immune phenotype.

Suggested Control Mice
C57BL/6
MMRRC Genetic QC Summary
The MMRRC Centers have developed a genetic QC pipeline using MiniMUGA array genotyping to provide additional information on strain backgrounds for MMRRC congenic and inbred strains. For more information on when data may be available, or to request genotyping for a strain of interest, please contact mmrrc@ucdavis.edu. Older strains may not have this information.
  • Immunology and Inflammation
Donor
Peggy S. Zelenka, Ph.D., National Eye Institute, National Institute of Health.
Michael Lenardo, National Institute of Allergy and Infectious Diseases.
Pushpa Pandiyan, Ph.D., Case Western Reserve University.

Colony and Husbandry Information

Colony Surveillance Program and Current Health Reports

Mice recovered from a cryo-archive will have health surveillance performed on recipient females. Health reports will be provided prior to shipment. If you require additional health status information, please email mmrrc@ucdavis.edu.
The MMRRC at UC Davis performed microbita analysis on feces samples received from the donating investigator while the mice were still at their facility. There is a MMRRC Fecal Microbiota report available for this strain.If you are interested in having the MMRRC at UC Davis perform microbiota analysis once the line is established at your facility please contact mmrrc@ucdavis.edu.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
N12
NOTE: "Hemizygote" as used here refers to males carrying a mutation on the X Chromosome or mice of either sex carrying an inserted transgene with no homologous allele on the other chromosome.
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Hetero/Hemizygotes are fertile: Yes Yes
Age Reproductive Decline: 4 to 6 months 4 to 6 months
Average litter size
4-6
Recommended wean age
3 Weeks
Average Pups Weaned
4-6

Order Request Information

Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

Cryopreserved material may be available upon request, please inquire to mmrrc@ucdavis.edu for more information.

The donor or their institution limits the distribution to non-profit institutions only.

Distribution of this strain requires submission of the MMRRC Conditions of Use (COU). A link to the COU web form will be provided via email after an order has been placed; the form should be completed then or the email forwarded to your institutional official for completion.

Additional charges may apply for any special requests. Shipping costs are in addition to the basic distribution/resuscitation fees. Information on shipping costs and any additional charges will be provided by the supplying MMRRC facility.

Click button to Request this one strain. (Use the MMRRC Catalog Search to request more than one strain.)
MMRRC Item # Description Distribution Fee / Unit (US $)
*Shipping & Handling not included*
Units Notes
044013-UCD-EMBRYO Cryo-preserved embryos $1,038.00 / Non-Profit Aliquot Approximate quantity2 : 20-40 embryos / aliquot
044013-UCD-SPERM Cryo-preserved spermatozoa $546.25 / Non-Profit Aliquot Approximate quantity3
044013-UCD-RESUS Litter recovered from cryo-archive $4,044.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.

1 The distribution fee covers the expense of rederiving mice from a live mouse; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

2 An aliquot contains a sufficient number of embryos (in one or more vials or straws and based on the transfer success rate of the MMRRC facility) to transfer into one to three recipients. The MMRRC makes no guarantee concerning embryo transfer success experienced in the recipient investigator's laboratory. Neither gender nor genotype ratios are guaranteed.

3 An aliquot is one straw or vial with sufficient sperm to recover at least one litter of mice, as per provided protocols, when performed at the MMRRC facility. The MMRRC makes no guarantee concerning the success of these procedures when performed outside the MMRRC facilities.

4 The distribution fee covers the expense of resuscitating mice from the cryo-archive; you will receive the resulting litter. The litter will contain at minimum one mutant carrier; the actual number of animals and the gender and genotype ratios will vary. (Typically, multiple breeder pairs can be established from the recovered litter.) Prior to shipment, the MMRRC will provide information about the animals recovered. If you anticipate or find that you need to request specific genotypes, genders or quantities of mice in excess of what is likely from a resuscitated litter, you may discuss available options and pricing with the supplying MMRRC facility.

To request material from the MMRRC: Please fill out our on-line request form (accessible from the catalog search results page, or click the Request this Strain button in the fees section). If you have questions or need assistance completing this form, you may call Customer Service at (800) 910-2291 (in USA or Canada) or (530) 757-5710 (international calls). Before you call, please have with you: the MMRRC item number, quantity needed, Bill-to and Ship-to contact information.