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Strain Detail Sheet

Strain Name:
B6J.Cg-Tg(Ube3a)2Bdph/Mmnc
Stock Number:
072143-UNC
Citation ID:
RRID:MMRRC_072143-UNC
Other Names:
Line C Ube3a+1

Strain Information

Gene Details

Ube3a
Name: ubiquitin protein ligase E3A
Synonyms: Hpve6a, E6-AP ubiquitin protein ligase, A130086L21Rik, 5830462N02Rik
Type: Gene
Species: Mouse
Chromosome: 7
NCBI: 22215
Homologene: 7988

Additional Resources
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ClinGen
Tg(Ube3a)2Bdph
Name: transgene insertion 2, Benjamin Philpot
Synonyms: Floxed-Ube3aOE
Type: Transgene
Species: Mus musculus
Chromosome: Unknown
Alteration at locus: Transgenic
Genetic Alterations
The bacterial artificial chromosome (BAC) clone RP24-178G7 contains the entire ubiquitin protein ligase E3A locus (Ube3a) as well as ~63 kbp of 5' flanking sequence and ~21 kbp of 3' flanking sequence. DNA sequences encoding a floxed Ube3a exon 7 (as per Ube3atm1.1Bdph) were introduced to the BAC via homologous recombination/BAC recombineering, replacing the native Ube3a exon 7 and surrounding sequences and resulting in a conditional transgenic allele. A single FRT site remains just upstream of floxed Ube3a exon 7. The transgenic insertion in the founder line was determined to comprise two tandem copies of the Tg(Ube3a)2Bdph transgene. Crossing to FLPo deleter mice resulted in efficient recombination of the FRT sites, enabling the production of this transgenic subline (i.e., Line C Ube3a+1), which harbors a single Tg(Ube3a)2Bdph copy. Ube3a expression was confirmed by qPCR, Western blot analysis, and immunofluorescent staining.

Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
    • ES Cell Line
    Not applicable

    Strain Description

    Phenotype

    This mouse strain overexpresses a single BAC transgenic copy of the mouse Ube3a gene (in addition to the two endogenous Ube3a gene copies) and thus models the level of UBE3A gene overdosage observed in individuals with interstitial duplications of the 15q11-q13 chromosomal region (Dup15q syndrome). Furthermore, this mouse strain exhibits aberrant forced swim behavior (reduced floating time) relative to wild-type littermate controls and significantly increased seizure mortality, reminiscent of the sudden unexpected death in epilepsy (SUDEP) phenotype in Dup15q syndrome.

    Conditional phenotype: Yes, these mice harbor a Ube3a transgene in which the exon 7 is flanked by LoxP sites. Cre-mediated excision of this critical Ube3a exon terminates transgenic Ube3a overexpression.

    Strain Development
    Generation of Ube3a overexpression (Ube3aOE) lines. A Ube3atm1a_KOMP_W (Ube3aKO1st) targeting cassette (KOMP) was recombineered into the 162 kb Ube3a BAC clone RP24-178G7 (pTARBAC1 vector backbone). Successfully recombineered BAC clones were identified by PCR and transformed into bacteria with inducible FLP recombinase expression to ultimately generate the conditional Ube3aOE BAC construct. Ube3aOE BAC was purified and integrity was confirmed using pulse-field electrophoresis. Purified Ube3aOE BAC was linearized via PI-SceI restriction digest prior to pronuclear microinjection of C57BL/6J fertilized eggs, which were subsequently implanted into pseudopregnant female mice. Six independent conditional Ube3aOE founder offspring (named A–F) were confirmed by PCR analysis of tail DNA. Two parental conditional Ube3aOE lines, C and E, were identified for use in this study following verification of germline transmission, mapping of chromosomal integration, assessment of transgene copy number via ddPCR, and confirmation of transgenic Ube3a expression. Both parental lines were confirmed to express two transgene copies (Ube3a+2). Line C Ube3a+2 mice was subsequently crossed to highly efficient FLPo deleter mice (RRID:IMSR_JAX:011065) to recombine the remaining FRT sites and collapse transgenic loci to a single conditional Ube3aOE copy, thereby establishing stable line C Ube3a+1 upon breeding out the FLPo allele in the subsequent generation. These lines have been maintained on a congenic C57BL/6J background.
    Suggested Control Mice
    C57BL/6J mice, preferably littermates, are appropriate controls for this strain.

    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

    • Neurobiology

    Strain Origin

    Donor
    Benjamin Philpot, Ph.D., The University of North Carolina- Chapel Hill.
    Primary Reference

    Punt AM, Judson MC, Sidorov MS, Williams BN, Johnson NS, Belder S, den Hertog D, Davis CR, Feygin MS, Lang PF, Jolfaei MA, Curran PJ, van IJcken WF, Elgersma Y, Philpot BD. Molecular and behavioral consequences of Ube3a gene overdosage in mice. JCI Insight. 2022 Sep 22;7(18):e158953. doi: 10.1172/jci.insight.158953. (Medline PMCID: 9675564)

    Colony and Husbandry Information

    Special Considerations

    No

    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
    >7
    Overall Breeding Performance
    Good

    Reproductive Statistics

    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: Undetermined Yes
    Hetero/Hemizygotes are fertile: Yes Yes
    Age Reproductive Decline: 7 to 9 months 7 to 9 months
    Average litter size
    7 to 9
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    7 to 9

    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.

    A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Carmen Melvin.

    Conditions of Distribution

    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.

    A Commercial License Agreement from the Donor is required for for-profit entities to use this strain. For more information, please contact Carmen Melvin

    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
    072143-UNC-RESUS Litter recovered from cryo-archive $3,242.00 / $5,382.00
    Non-Profit / For-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.