Strain Detail Sheet

Strain Name:
C57BL/6J-Il17fem1Gaff/Mmnc
Stock Number:
068083-UNC
Citation ID:
RRID:MMRRC_068083-UNC
Other Names:
Il17f.S65L

Strain Information

Gene Details

Il17fem1Gaff
Name: interleukin 17F; endonuclease-mediated mutation 1, Sarah Gaffen
Synonyms: Il17fS65L
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 1
Alteration at locus: CRISPR
Il17f
Name: interleukin 17F
Type: Gene
Species: Mouse
Chromosome: 1
Alteration at locus: CRISPR
NCBI: 257630
Homologene: 17115
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Genetic Alterations
The mice contain a mutation predicting a serine-to-leucine substitution at codon 93. This site is conserved in the human IL17F gene, and the introduced alteration was based on a naturally-occurring mutation found in a human family that caused susceptibility to mucosal candidiasis. The associated publication refers to this as S65L, in reference to the mature protein.

HGVS nomenclature:

  • Genbank RefSeq - mRNA: NM_145856.2
  • Genbank RefSeq, protein: NP_665855.2
  • Variant, nucleic acid level: c.276_282delinsTCTAGAA
  • Variant, amino acid level, predicted: p.Ser93Leu
    Check this variant: LUMC Mutalyzer
    Note: This variant is described in the literature as S65L, which uses positional referencing to the mature protein (i.e., absence of the secretory signal sequence).

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
    Mice carry a mutation in the Il17f cytokine gene. In the homozygous state, the animals exhibit increased susceptibility to oral Candida albicans infections. At baseline, the mice appear healthy and are fertile.
    Strain Development
    The line was created by CRISPR/Cas9 in the Transgenic and Gene Targeting (TGT) and Innovative Technologies Development (ITD) Core facilities in the Department of Immunology, University of Pittsburgh. Briefly, a Cas9 target sequence overlapping the Ser65 codon in the mature Il17f sequence (following signal peptide cleavage) was selected. The intended substitution converts Serine65 to leucine, which contemporaneously disrupts an Hpy188I restriction site to facilitate genotyping. The sequence also contains a silent substitution, bringing a total of 4 mismatches between the sgRNA target sequence and the designed allele, thus limiting further re-editing of the mutant allele by Cas9.

    C57BL/6J embryos were microinjected with the sgRNA, the HDR oligonucleotide, and Cas9 mRNA. Embryos that developed to the 2-cell stage were transferred into pseudopregnant female surrogates. Sixteen founders were identified by PCR amplification of the target region followed by restriction digestion of the PCR product with Hpy188I. The correct sequence was validated by Sanger sequencing of the PCR products following TOPO cloning. The expanded line was backcrossed twice to C57BL/6J mice prior to colony expansion.
    Suggested Control Mice
    C57BL/6J

    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

    • Immunology and Inflammation

    Strain Origin

    Donor
    Sarah Gaffen, Ph.D., University of Pittsburgh.
    Primary Reference

    Zhou C, Monin L, Gordon R, Aggor FEY, Bechara R, Edwards TN, Kaplan DH, Gingras S, Gaffen SL. An IL-17F.S65L Knock-In Mouse Reveals Similarities and Differences in IL-17F Function in Oral Candidiasis: A New Tool to Understand IL-17F. J Immunol. 2020 Aug 1;205(3):720-730. doi: 10.4049/jimmunol.2000394. Epub 2020 Jun 29. (Medline PMID: 32601099)

    Colony and Husbandry Information

    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
    Sib-mating
    Generation
    N2 (C57BL/6J)
    Overall Breeding Performance
    Excellent

    Reproductive Statistics

    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
    Average litter size
    4 to 6
    Recommended wean age
    3 Weeks
    Average Pups Weaned
    4 to 6

    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 Stefanie Frelick.

    Conditions of Distribution

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

    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.

    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
    068083-UNC-SPERM Cryo-preserved spermatozoa $564.00 / $937.00
    Non-Profit / For-Profit    		 Aliquot Approximate quantity3
    068083-UNC-RESUS Litter recovered from cryo-archive $2,914.00 / $4,837.00
    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.