Strain Name:
B6N.129S6-Gapdhstm1Dao/Mmnc
Stock Number:
036555-UNC
Citation ID:
RRID:MMRRC_036555-UNC
Other Names:
Gapdhs -/-, Gapds -/-

Strain Information

Gapdhstm1Dao
Name: glyceraldehyde-3-phosphate dehydrogenase, spermatogenic; targeted mutation 1, Deborah A O'Brien
Synonyms: Gapds-
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 7
Alteration at locus: Knockout
Gapdhs
Name: glyceraldehyde-3-phosphate dehydrogenase, spermatogenic
Synonyms: Gapds, Gapd-s
Type: Gene
Species: Mouse
Chromosome: 7
Alteration at locus: Knockout
NCBI: 14447
Homologene: 101265
Genetic Alterations
Gapdhs was isolated from a P1 phage library of 129/OlaHsd mouse genomic DNA. Nucleotide numbers in the Gapdhs genomic sequence refer to GenBank accession no. U09964. After digestion with Tth111I and SspI, the DNA restriction fragment beginning in exon 1 and ending in intron 4 (nucleotides 1887–7193) was cloned into the SrfI site of the cloning vector pUCBM21/KO. This fragment, along with thymidine kinase (tk) and neomycin resistance (neo) genes, was ligated into the NotI–XhoI site of pMC1TKbpA, which contains another tk cassette. The Gapdhs DNA fragment beginning in exon 6 and ending in exon 9 (nucleotides 8108–9181) was amplified by PCR using the P1 clone as template and then ligated into the SpeI–XbaI site of pMC1TKbpA to produce the gene-targeting construct pL3KaSTK2. Transfection of pL3KaSTK2 DNA, screening of targeted TC-1 embryonic stem (ES) cells, and blastocyst injections were performed. The PCR primers used to detect homologous recombination were forward 5-AGCGTTGGCTACCCGTGATA-3 and reverse 5'-CGTGATAGCCGAGTAAGAAGCAGG-3, corresponding to sequences in the neo gene and exon 9, respectively.
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
  • ES Cell Line
    TC1/TC-1 derived from 129S6/SvEvTac
    Phenotype
    Homozygous: Gapdhs -/- males are infertile, although they produced comparable numbers of sperm to those of wild-type males and have normal testis weights, body weights, and testis histology. Mating behavior also appears normal when assessed by housing individual Gapdhs-/- males with two superovulated females for 2 h. However, Gapdhs -/- males sired no pups when mated continuously with two wild-type females for 1 month, followed by a second month with two different females (n =7 males, n =28 females). Sperm from Gapdhs -/- males have extremely low ATP levels and profound defects in motility, with little or no forward movement. Gapdhs-/- females had normal fertility. Gapdhs -/- mice of both sexes were produced in the expected proportions from heterozygous matings (64 homozygous mutant, 140 heterozygous, 59 wild-type) and were not visibly different from their wild-type littermates.
    Heterozygous: Gapdhs+/- males and females have normal fertility.
    Mammalian Phenotype Terms
    Allelic Composition: Gapdhstm1Dao/Gapdhstm1Dao (Genetic Background: involves: 129S6/SvEvTac * C57BL/6N )

    Strain Development

    Chimeric males generated from correctly targeted ES cells were mated with C57BL/6N females to obtain germ-line transmission. Heterozygous animals were mated to produce Gapdhs-/- and Gapdhs+/- males for the analysis of fertility and sperm function. Heterozygous males from each generation were backcrossed to C57BL/6N females for ten generations.

    A sample from this MMRRC strain was analyzed using the Mouse Universal Genotyping Array (MUGA) and MMRRC computational tools were used to assess the genetic background. A summary of the data can be found here.

    MMRRC Strain Reference Strains Genomic makeup (Mb)
    Strain Name MMRRC ID # Reference 1 Reference 2 Ref 1 Ref 2 Het Unknown
    B6N.129S6-Gapdhstm1Dao/Mmnc 36555 C57BL/6NJ 129S6/SvEvTac 2574 0 10 0
    Suggested Control Mice
    Wild-type littermates or C57BL/6N mice
    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.
    • Cell Biology
    • Developmental Biology
    • Metabolism
    • Models for Human Disease
    • Reproduction
    Donor
    Deborah A O'Brien, Ph.D., University of North Carolina at Chapel Hill School of Medicine.

    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
    MMRRC Breeding System
    Backcross or Sib-mating
    Breeding Scheme(s)

    C57BL/6N female x Heterozygous male

    Heterozygous female x Heterozygous male littermates

    Generation
    N10
    Overall Breeding Performance
    Good
    Viability and Fertility: Female Male Comments
    Homozygotes are viable: Yes Yes
    Homozygotes are fertile: Yes No
    Heterozygotes are fertile: Yes Yes
    Age Reproductive Decline: 8 months 10 months
    Average litter size
    5-9
    Recommended wean age
    3 weeks

    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.

    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.

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

    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
    036555-UNC-SPERM Cryo-preserved spermatozoa $564.00 / Non-Profit Aliquot Approximate quantity3
    036555-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.