Strain Name:
B6J.129X1-Gt(ROSA)26Sortm1(CAG-Vps4b*)1Hjug/Mmnc
Stock Number:
046320-UNC
Citation ID:
RRID:MMRRC_046320-UNC
Other Names:
Gt(ROSA)26Sor tm1(LSL mVPS4B EQ)/Hjug

Strain Information

CAG
Name: CMV early enhancer/chicken beta actin promoter/rabbit beta globin splice acceptor
Type: DNA Segment
Species: Multi-species
Chromosome:
Gt(ROSA)26Sor
Name: gene trap ROSA 26, Philippe Soriano
Synonyms: beta geo, R26, Gtrosa26, Thumpd3as1, Gtrgeo26, ROSA26
Type: Gene
Species: Mus musculus (mouse)
Chromosome: 6
NCBI: 14910
Vps4b
Name: vacuolar protein sorting 4B
Synonyms: Skd1, 8030489C12Rik
Type: Gene
Species: Mouse
Chromosome: 1
NCBI: 20479
Homologene: 37976
Genetic Alterations
A lox-STOP-lox cassette followed by the ORF of murine Vps4b was targeted to the Rosa26 locus. In order to create a dominant-negative Vps4b allele, a point mutation was introduced as follows:

Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
  • ES Cell Line
    EC7.1
    Phenotype
    The non-recombined allele has no phenotype. Postnatal endothelial cell-specific recombination results in CNS angiogenesis defects, blood-CNS-barrier defects, and lethality within 4-6 days.
    Strain Development
    Cre-activatable Vps4b E235Q (Vps4bEQ) mice were generated by a knock-in strategy into the Rosa26 locus. Mouse Vps4bEQ cDNA with MycHis tag and bovine growth hormone polyadenylation sequence was amplified from pEGFP-N1-mVps4b-EQ-MycHis plasmid (gift from Dr. Phyllis Hanson) and subcloned into NotI/EcoRI sites of transfer vector pMAT1. The Vps4bEQ sequence together with pMAT1-encoded portions of the CAG promoter and lox-STOP-lox sequence were subcloned into the MreI and EcoRI sites of pHW249 targeting vector (pMAT1 and pHW249 were kindly provided by Dr. Jeremy Nathans). The targeting construct was linearized with KpnI before electroporation into EC7.1 ES cells in the University of Colorado Anschutz Medical Campus gene targeting core. Positive ES cell clones were identified by PCR using primers upstream and downstream of the 5’ arm (5’- AAGAAGAGGCTGTGCTTTG-3’ / 5’- TTGGCGTTACTATGGGAAC-3’) as well as using primers upstream and downstream of the 3’ arm (5’-GAACCAGCTGGGGCTCGACTAGAG-3’ / 5’-CAAGCACTGTCCTGTCCTCAAG-3’). Selected positive clones were karyotyped and two correct clones were injected into blastocyst to generate chimeric mice. Chimeric mice were crossed with C57BL/6J mice and genotyped for the presence of a 483-bp Vps4b amplicon using the primers 5’-TGGAGATAAACTCTTGGAGCC-3’ / 5’-CCGGAATTCTCCGCCTCAGAAGCCATAGAGC-3’.
    Suggested Control Mice
    wild type, R26 LSL mVps4b EQ; CreERT2-negative with tamoxifen, R26 LSL mVps4b EQ; CreERT2-positive without tamoxifen
    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.
    Donor
    Harald Junge, Ph.D University of Colorado at Boulder
    Primary Reference

    Zhang C, Lai MB, Khandan L, Lee LA, Chen Z, Junge HJ. Norrin-induced Frizzled4endocytosis and endo-lysosomal trafficking control retinal angiogenesis andbarrier function. Nat Commun. 2017 Jul 4;8:16050. doi: 10.1038/ncomms16050.(Medline PMID: 28675177)

    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_health@med.unc.edu.
    Coat Color
    Black
    Eye
    Black
    MMRRC Breeding System
    Backcross
    Generation
    N7
    Overall Breeding Performance
    Good
    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.
    Average litter size
    Undetermined
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    Undetermined

    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@med.unc.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
    046320-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.