Loading...

Strain Detail Sheet

Strain Name:
129S6(Cg)-Krastm5.1Cmco/Mmnc
Stock Number:
071549-UNC
Citation ID:
RRID:MMRRC_071549-UNC
Other Names:
Kras, Kras-LSL-comG12D

Strain Information

Gene Details

Krastm5.1Cmco
Name: Kirsten rat sarcoma viral oncogene homolog; targeted mutation 5.1, Christopher M Counter
Synonyms: KrasLSL-comG12D
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 6
Alteration at locus: Conditional
Kras
Name: Kirsten rat sarcoma viral oncogene homolog
Synonyms: Ki-ras, Kras-2, K-ras, Kras2
Type: Gene
Species: Mouse
Chromosome: 6
Alteration at locus: Conditional
NCBI: 16653
HGNC: HGNC:6407
Homologene: 37990

Additional Resources
IMPC
Google
Entrez
PubMed
IMSR
Gene Ontology
BioGPS
Gene Cloud
IGTC
Mouse Phenome DB
Mouse Mine
KOMP Phenotyping
Monarch
GeneCards
ClinGen
Genetic Alterations
c
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
    • ES Cell Line
    G4 derived from (129S6/SvEvTac x C57BL/6Ncr)F1

    Strain Description

    Phenotype
    Considering homozygous mutant Kras alleles are embryonically lethal, the strain was kept heterozygous under 129 background. Phenotypes such as tumors of hematopietic system, lung, oral epithelia, and forestomach epithelia were observed.
    • Increased endogenous expression
    • Median survival was around 51 days.
    • Activating only in the lung using the CC10-CreER background revealed tumors starting 1 month after tamoxifen injection.
    • Conditional phenotype: Yes, LSL cassette can be recombined to activate the allele by a ubiquitous CRE driver (Rosa26CreERT2) and H&E staining performed of various tissues.
    MeSH Terms
    • Animals
    • Carcinogenesis/genetics
    • Cell Transformation, Neoplastic/genetics
    • Genes, ras
    • Mice
    • Mutation
    • Neoplasms/genetics
    • Proto-Oncogene Proteins p21(ras)/genetics
    • Proto-Oncogene Proteins p21(ras)/metabolism
    Strain Development
    A bacteria artificial chromosome was engineered with 7.5 kbp of 5′ flanking sequence Kras intron 1 DNA, a Lox-STOP-Lox cassette (LSL), the first three coding exons fused together and encoded by the most commonly used codons in the mouse genome (com) with a G12D oncogenic mutation, followed by the N-terminal 564 bp of intron 4, an FRT-Neomycin-FRT cassette, and a further 1.5 kbp of 3′ flanking sequence. The targeting cDNAs were each cloned into the targeting PL253 vector and electroporated into G4 ES cells.

    After selection, clones were screened using PCR and positive clones were selected for each engineered Kras allele, expanded, and frozen. At least two targeted clones per allele confirmed with a Southern hybridization were then microinjected into blastocysts to produce chimeras. KrasLSL-comG12D(+neo)/+ chimeras (still retaining the neo cassette in the engineered Kras alleles) were crossed back to 129S6 mice. A genotyping PCR specific to the engineered Kras alleles with neo cassette was used to screen for germline transmission in clones. 129S6-KrasLSL-comG12D(+neo)/+ cohort was crossed with ACTBFLPe/FLPe (Jackson Laboratory, strain 003800) mice to remove the selection marker via FLP-mediated excision of the neo cassette (Dymecki, 1996). Removal of the neo cassette was confirmed with genotyping PCR. Resultant 129S6-KrasLSL-comG12D(-neo)/+ were backcrossed with 129S6 for five consecutive generations and maintained in the 129 background.
    Suggested Control Mice
    129

    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

    • Cancer
    • Models for Human Disease

    Strain Origin

    Donor
    Christopher Counter, Ph.D., Duke University.
    Primary Reference

    Le Roux Ö, Pershing NLK, Kaltenbrun E, Newman NJ, Everitt JI, Baldelli E, Pierobon M, Petricoin EF, Counter CM. Genetically manipulating endogenous Kras levels and oncogenic mutations in vivo influences tissue patterning of murine tumorigenesis. Elife. 2022 Sep 7;11:e75715. doi: 10.7554/eLife.75715. (Medline PMID: 36069770)

    Colony and Husbandry Information

    Special Considerations

    When maintained as a live colony, heterozygotes may be bred. Homozygotes are embryonic lethal.

    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
    Agouti
    Eye
    Black

    Breeding

    MMRRC Breeding System
    Backcross
    Generation
    10+
    Overall Breeding Performance
    Good

    Reproductive Statistics

    Viability and Fertility: Female Male Comments
    Homozygotes are viable: No No
    Homozygotes are fertile: N/A N/A
    Heterozygotes are fertile: Yes Yes
    Age Reproductive Decline: 10 to 12 months Greater than 12 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 Shweta Krishnan.

    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 Shweta Krishnan

    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
    071549-UNC-RESUS Litter recovered from cryo-archive $3,088.00 / $5,126.00
    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.