Strain Detail Sheet

Strain Name:
B6J.Cg-Tspan12tm1.1Hjug/Mmnc
Stock Number:
046319-UNC
Citation ID:
RRID:MMRRC_046319-UNC
Other Names:
Tspan12tm1.1Hjug

Strain Information

Gene Details

Tspan12tm1.1Hjug
Name: tetraspanin 12; targeted mutation 1.1, Harald Junge
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 6
Alteration at locus: Knockout
Tspan12
Name: tetraspanin 12
Synonyms: Tm4sf12
Type: Gene
Species: Mouse
Chromosome: 6
Alteration at locus: Knockout
NCBI: 269831
Homologene: 8212
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Genetic Alterations
A LoxP flanked allele, in which the critical exon 3 (containing the start codon and major portions of the first transmembrane segment) can be removed by Cre-mediated recombination.
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
    • ES Cell Line
    EC7.1

    Strain Description

    Phenotype
    Homozygous mice carrying the non-recombined floxed allele have no phenotype. Following expression of cre, postnatal endothelial cell-specific recombination of Tspan12tm1.1Hjug causes lack of intraretinal capillaries and blood-retina barrier defects. Endothelial cell-specific loss of TSPAN12 in mature mice strongly impairs blood-retina barrier maintenance without affecting vascular morphogenesis, pericyte coverage, or perfusion. Long-term BRB defects are associated with immunoglobulin extravasation, complement deposition, cystoid edema, and impaired b-wave in electroretinograms.
    Strain Development
    The targeting construct for generating a Tspan12 floxed allele, pTspan12-LFNFL, contained the following sequence segments: exon3 (containing the start codon and major portions of the first transmembrane segment) was flanked on the 3’ end by a loxP site, and on the 5’ end by a FRT-flanked PGK-neo-bGH cassette (for positive selection of ES cells) and the 5’ loxP site. The 5’ homology arm (left arm) consisted of 2978 bp sequence containing exon1, exon2, and adjacent sequences. The 3’ homologous arm (right arm) consisted of 2510 bp of intronic sequences downstream of exon3. The construct contained a thymidine kinase cassette outside the homology arms for negative selection of ES cells. The linearized construct was electroporated into EC7.1 ES cells and subjected to negative/positive selection with ganciclovir and G418, respectively. ES cell clones were screened using genomic PCRs that amplified the right and left arms separately, using primers outside the left or outside the right homology arms, each combined with a primer inside the targeting cassette. In addition, the entire targeted region was amplified using primers outside each homology arm. PCR products were further analyzed by restriction digest. A karyotyped ES cell clone (D11) was injected into C57BL/6 blastocysts and chimeras with germline transmission were identified by PCR. The FRT flanked PGKneo cassette was removed by Flp recombination in the germline using FLPo mice (B6.Cg-Tg(Pgk1-flpo)10Sykr/J - JAX Strain #011065), and recombined offspring were identified by PCR. The floxed conditional allele was genotyped using mTSPAN12_LA_fw oligo (5’-GTCTCAAGGGCCATGCCACTG-3’) and TSP12ex3-rev oligo (5’- CCACATTCTACTGGTCCAGGCAGAG-3’), yielding a 403 bp amplicon on the floxed allele and a 301 bp amplicon on the wild-type allele. Mice were backcrossed on a C57BL/6J background.
    Suggested Control Mice
    wild-type, Tspan12flox/flox;Cre-negative, Tspan12+/+;Cre-positive

    MMRRC Genetic QC

    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@med.unc.edu. Older strains may not have this information.

    Research Applications

    • Cancer
    • Cardiovascular
    • Developmental Biology
    • Models for Human Disease
    • Neurobiology

    Strain Origin

    Donor
    Harald Junge, Ph.D University of Colorado at Boulder
    Primary Reference
    (unpublished)

    Colony and Husbandry Information

    Health Status Report

    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_health@med.unc.edu.

    Appearance

    Coat Color
    Black
    Eye
    Black

    Breeding

    MMRRC Breeding System
    Backcross
    Generation
    N10
    Overall Breeding Performance
    Good

    Reproductive Statistics

    Average litter size
    Undetermined
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    Undetermined

    Order Request Information

    Availability Level

    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@med.unc.edu for more information.

    Conditions of Distribution

    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.

    Fees

    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
    046319-UNC-RESUS Litter recovered from cryo-archive $2,914.00 / Non-Profit Litter Recovered litter4; additional fees for any special requests.
    Cryopreserved material may be available upon request, please inquire to mmrrc@med.unc.edu for more information.

    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.