Strain Name:
STOCK Tg(Astl/mCherry)1Dean/Mmnc
Stock Number:
066521-UNC
Citation ID:
RRID:MMRRC_066521-UNC
Other Names:
Astl-mCherry

Strain Information

Astl
Name: astacin like metalloendopeptidase
Synonyms: Ovastacin, C87576, Sas1b
Type: Gene
Species: Mouse
Chromosome: 2
NCBI: 215095
Homologene: 128191
mCherry
Name: red fluorescent fluorophore, mCherry
Type: Gene
Species: Discosoma sp. (mushroom coral)
Chromosome:
Tg(Astl/mCherry)1Dean
Name: transgene insertion 1, Jurrien Dean
Synonyms: AstlmCherry, Tg(Astl-mCherry)1Dean, ovastacinmCherry
Type: Transgene
Species: Mus musculus (mouse)
Chromosome:
Alteration at locus: Transgenic (random, gene disruption)
Genetic Alterations
This transgenic mouse line has a mCherry reporter fused with the last codon of Astl gene, resulting in an Astl-mCherry transgene.
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
  • Phenotype
    Eggs of mice produce red cortical granules.


    Astl-mCherry transgenic mice provide an authentic biological marker for cortical granules. The zona pellucida and ovastacin proteins transverse the endomembrane system during oogenesis during which passage ZP2 remains intact. Secreted ZP2 is incorporated into the extracellular zona pellucida surrounding ovulated eggs and is cleaved in the zona matrix surrounding 2-cell embryos. In addition to a rapid, cortical-granule independent, post-fertilization block to gamete fusion, ZP2 cleavage provides a definitive block to polyspermy in that sperm that do not bind cannot penetrate the zona matrix nor fuse with the egg plasma membrane. The single-copy Astl gene encodes ovastacin (435 aa), an oocyte-specific metalloendoprotease that has been located by immunohistochemistry in cortical granules at the periphery of growing oocytes and ovulated eggs.

    MeSH Terms
    • Animals
    • Cytoplasmic Granules/metabolism
    • Female
    • Fertilization
    • Metalloproteases/chemistry
    • Metalloproteases/genetics
    • Metalloproteases/metabolism
    • Mice
    • Oocytes/metabolism
    • Protein Sorting Signals
    • Protein Transport
    • Proteolysis
    • Zona Pellucida Glycoproteins/metabolism
    Strain Development
    To construct the transgene, bacterial artificial chromosome (BAC) DNA that included mouse Astl (BMQ56H22) was transformed into SW102 bacterial cells containing the λ-prophage recombineering system. A PCR fragment (1331 bp) containing the galK operon flanked by 50-bp homologous to the Astl gene that would insert the galK gene at the stop codon in the 3’ region of the gene was amplified using Long Amp Taq Polymerase. After digestion with DpnI and overnight gel purification, the PCR fragment was electroporated into SW102 cells containing the BAC, and recombinants were selected by growth on minimal media with galactose. Using a clone from this first step, the galK cassette was replaced by recombineering with a second PCR fragment (880 bp) encoding mCherry with 50-bp homology arms. Clones were selected on minimal media with 2-deoxy-galactose and confirmed by DNA sequence. Finally, a NotI fragment containing the Astl-mCherry transgene was retrieved from the BAC with pl253, and the fidelity of coding regions was confirmed by DNA sequence. After gel purification, the Astl-mCherry transgene was injected into the male pronucleus of >200 one-cell zygotes (FVB/N) which were transferred to pseudopregnant female mice. Offspring were genotyped using tail DNA and Astl exon 9 oligonucleotides that distinguished between the normal allele (268 bp) and the Astl-mCherry transgene (967 bp). From 30 pups, 6 founders were identified that passed the transgene through the germline. Three lines were maintained; the line described here was additionally crossed to B6D2F1 without further backcrossing.
    Suggested Control Mice
    Wild-type
    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
    Jurrien Dean, M.D., National Institutes of Health (NIH), NIDDK.
    Primary Reference

    Xiong B, Zhao Y, Beall S, Sadusky AB, Dean J. A Unique Egg Cortical GranuleLocalization Motif Is Required for Ovastacin Sequestration to Prevent PrematureZP2 Cleavage and Ensure Female Fertility in Mice. PLoS Genet. 2017 Jan23;13(1):e1006580. doi: 10.1371/journal.pgen.1006580. eCollection 2017 Jan.(Medline PMID: 28114310)

    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
    Pink
    MMRRC Breeding System
    Random intra-strain mating
    Generation
    F12?
    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.
    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 to 6
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    4 to 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@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
    066521-UNC-SPERM Cryo-preserved spermatozoa $564.00 / Non-Profit Aliquot Approximate quantity3
    066521-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.

    Some MMRRC strains were submitted by the donor as cryopreserved germplasm, and because these strains were not cryopreserved by the MMRRC, we have not assessed the quality, nor genotype, of this material. Our expertise in reviving mice from various qualities of frozen sperm and embryos, using methods like IVF and ICSI, gives us confidence in successful recoveries in most cases. However, due to the uncertain quality of these samples, we'll limit revival attempts to two per order. Additional attempts are available for a fee, on top of standard charges, if requested. It's important to note that some strains may lack the expected mutation, so we can't assure successful order fulfillment until we attempt to revive the strain.

    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.