Strain Name:
B6N.Cg-Ppifem1Mure/Mmnc
Stock Number:
066982-UNC
Citation ID:
RRID:MMRRC_066982-UNC
Other Names:
CypD C202S

Strain Information

Ppifem1Mure
Name: peptidylprolyl isomerase F (cyclophilin F); endonuclease-mediated mutation 1, Elizbeth Murphy
Synonyms: Ppifp.Cys202Ser
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 14
Alteration at locus: CRISPR
Ppif
Name: peptidylprolyl isomerase F (cyclophilin F)
Synonyms: mitochondrial Cyclophilin D, CyP-F, CypD, CyP-D
Type: Gene
Species: Mouse
Chromosome: 14
Alteration at locus: CRISPR
NCBI: 105675
HGNC: HGNC:9259
Homologene: 38696
Genetic Alterations
G-to-C mutation that predicts the replacement of cysteine 202 with a serine (C202S). The allele also has a three synonymous single-base exchanges (NM_134084.1:c.[609C>A;612G>A;613T>C]) to avoid Cas9 recognition of the inserted sgRNA.

HGVS nomenclature:

  • Genbank RefSeq - mRNA: NM_134084.1
  • Genbank RefSeq, protein: NP_598845.1
  • Variant, nucleic acid level: c.[605G>C;609C>A;612G>A;613T>C]
  • Variant, amino acid level, predicted: p.[Cys202Ser;p.=;p.=;p.=]
    Check this variant: LUMC Mutalyzer
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
  • ES Cell Line
    Not applicable
    Phenotype
    Homozygous: Protected against cardiac ischemia/reperfusion injury. Otherwise normal.Heterozygous: normal.

    Note from the Donating Investigator: "This mouse is a unique research tool to study redox modifications of Cysteine 202, which has been implicated in ischemia/reperfusion injury in vivo. Cyclophilin D (encoded by the gene Ppif) plays an important role in regulating cell death in the heart."
    Strain Development
    The strand guide RNA (sgRNA) was designed to bind to ~20 nucleotides within the Ppif (CypD) locus and was centered near the nucleotides that predict p.Cys202Ser (C202S). Oligo2 containing four mutations (see section Genetic Alteration) was injected with the CRISPR/Cas9 components to be incorporated into the genomic DNA via homologous recombination at the cut CypD locus. Recombination of Oligo2 produced a CypD locus with the aforementioned 4 exchanges, three silent and the fourth predicting a cysteine-to-serine substitution at codon 202. Zygotes from B6CBAF1/J were microinjected with CRISPR/Cas9, sgRNA and CypD Oligo2. Microinjected zygotes were cultured to the 2-cell stage and implanted into the oviducts of pseudopregnant foster mothers. PCR and Sanger sequencing were used to identify the genotype of the offspring. Founder mice were subsequently backcrossed with C57BL6/N to N11.
    Suggested Control Mice
    C57BL/6N
    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.
    • Apoptosis
    • Cardiovascular
    • Cell Biology
    • Research Tools
    Donor
    Elizabeth Murphy, Ph.D., National Institutes of Health, National Heart, Lung, and Blood Institute (NHLB).
    Primary Reference

    Amanakis G, Sun J, Fergusson MM, McGinty S, Liu C, Molkentin JD, Murphy E.Cysteine 202 of Cyclophilin D is a site of multiple post-translationalmodifications and plays a role in cardioprotection. Cardiovasc Res. 2020 Mar 4.pii: cvaa053. doi: 10.1093/cvr/cvaa053. [Epub ahead of print] (Medline PMID: 32129829)

    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
    N11 (C57BL/6N)
    Overall Breeding Performance
    Excellent
    Viability and Fertility: Female Male Comments
    Homozygotes are viable: Yes Yes
    Homozygotes are fertile: Yes Yes
    Heterozygotes are fertile: Yes Yes
    Age Reproductive Decline: Undetermined Undetermined
    Bred to Homozygosity
    Yes
    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
    066982-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.