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Strain Detail Sheet

Strain Name:
B6J.B6N-Myo10tm1a(KOMP)Wtsi/Mmnc
Stock Number:
071696-UNC
Citation ID:
RRID:MMRRC_071696-UNC
Other Names:
Myo10tm1a

Strain Information

Gene Details

Myo10tm1a(KOMP)Wtsi
Name: myosin X; targeted mutation 1a, Wellcome Trust Sanger Institute
Synonyms: Myo10tm1a
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 15
Alteration at locus: Targeted Mutation
Myo10
Name: myosin X
Synonyms: myosin-X, D15Ertd600e
Type: Gene
Species: Mouse
Chromosome: 15
Alteration at locus: Targeted Mutation
NCBI: 17909
HGNC: HGNC:7593
Homologene: 36328

Additional Resources
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Google
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Gene Ontology
BioGPS
Gene Cloud
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Mouse Phenome DB
Mouse Mine
KOMP Phenotyping
Monarch
GeneCards
ClinGen
Genetic Alterations
The L1L2_Bact_P cassette was inserted at position 25785401 of Chromosome 15 upstream of the critical exon(s) (Build GRCm39). The cassette is composed of an FRT site followed by lacZ sequence and a loxP site. This first loxP site is followed by a neomycin resistance gene under the control of the human beta-actin promoter, SV40 polyA, a second FRT site and a second loxP site. A third loxP site is inserted downstream of the targeted exon(s) at position 25786400. The critical exon(s) is/are thus flanked by loxP sites. A "conditional ready" (floxed) allele can be created by flp recombinase expression in mice carrying this allele. Subsequent cre expression results in a knockout mouse. If cre expression occurs without flp expression, a reporter knockout mouse will be created.
Genotype Determination
  • Genotyping Protocol(s)
  • Center protocol and contact for technical support will be shipped with mice.
    • ES Cell Line

    JM8A1.N3

    Strain Description

    Phenotype
    Approximately half of Myo10tm1a homozygotes exhibit the lethal neural tube defect of exencephaly. The tm1a homozygotes that are not affected by exencephaly exhibit 100% incidence of a white belly-spot and 100% incidence of persistent hyaloid vasculature. Numerous other eye defects are observed at lower frequencies including anophthalmia, microphthalmia, coloboma, and defects in lens formation. Approximately half of tm1a homozygotes exhibit syndactyly. Based on KOMP data, tm2 homozygotes also have white belly spots and a high incidence of persistent hyaloid vasculature. Based on the characterization reported in our manuscript on Myo10 null mice, the tm1a and m1j alleles exhibit a similar core phenotype, but the tm1a allele has conditional potential, making it much more useful, especially given that homozygous KOs exhibit a large number of defects and ~50% lethality. Homozygous: Born at less than half of the expected Mendelian ratio, which attributed largely to the high frequency (~50%) of exencephaly in the homozygotes. Homozygotes that are not exencephalic are able to develop into adult mice and breed. Homozygotes average ~20% lighter in weight than wild-type and nearly all homozygotes have a white belly spot although it is sometimes quite small. 100% of homozygotes whose eyes were dissected exhibited persistent fetal vasculature. Approximately 50% of Myo10tm1a homozygotes exhibited webbed digits. Hetero/Hemizygous: Heterozygotes appear normal and are not obviously different from wild-type.

    Conditional phenotype: The Myo10tm1a allele contains a KO first cassette inserted just upstream of exon 27 of full-length Myo10 (exons 1-41, aa 1-2062). The strong splice acceptor site, IRES, and B-gal encoded by the KO-first cassette are expected to disrupt full-length Myo10 (NM_019472.2) and the three headless Myo10 transcripts (XM_192774.2, XM_006520025.3, and XM_006520024.3) after the amino acid corresponding to residue 1190 of full-length Myo10 due to the presence of 61 amino acids and a stop codon from the KO-first cassette. The KO first cassette also contains FRT sites to allow FLP-mediated recombination to remove the KO first cassette and generate a tm1c conditional (cKO) allele. The loxP sites flanking exon 27 allow cre mediated deletion of exon 27, which allows conversion of the floxed tm1c allele into the tm1d deletion allele.

    Mammalian Phenotype Terms
    Allelic Composition: (Genetic Background: )
    MeSH Terms
    • Animals
    • Female
    • Male
    • Mice
    • Mice, Inbred C57BL
    • Mice, Knockout
    • Myosins/physiology
    • Neovascularization, Pathologic
    • Neural Tube/physiopathology
    • Ophthalmic Artery/metabolism
    • Ophthalmic Artery/physiopathology
    • Pigmentation
    • Pseudopodia/metabolism
    • Pseudopodia/pathology
    • Vitreous Body/blood supply
    • Vitreous Body/metabolism
    • Vitreous Body/pathology
    Strain Development
    The Myo10tm1a KO-first allele was generated from C57BL6EPD0332_3-B02 ES cells (see KOMP project 50217). These ES cells were produced from JM8A3.N1 cells, an ES line derived from C57BL/6N mice. The KOMP ES cells were injected into blastocysts and the resulting chimeras were bred to albino C57BL/6 mice to get germline transmission on a C57BL/6 background. The resulting germline mice were crossed with one another and with C57BL/6 mice prior to being sent to the Cheney lab at UNC. At Cheney lab, the original tm1a male heterozygote was bred to C57BL/6J (Jackson Strain #000664) and a colony was established by mating the tm1a offspring to one another. Some Myo10tm1a mice on this mixed C57BL/6J and C57BL/6N background were deposited with the MMRRC as strain #43822 while others were backcrossed to make them congenic on C57BL/6J, which was confirmed by MiniMuga testing in 2022. The mice were then maintained by intrastrain mating prior to being deposited to the MMRRC.
    Suggested Control Mice
    WT littermates, C57BL/6J. Approximately half of homozygous Myo10 KO embryos exhibit the lethal defect of exencephaly, it can be advantageous to increase the yield of homozygous KOs by breeding heterozygous males to homozygous females and using heterozygotes as controls.

    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
    • Cardiovascular
    • Cell Biology
    • Dermatology
    • Developmental Biology
    • Immunology and Inflammation
    • Internal/Organ
    • Models for Human Disease
    • Neurobiology
    • Reproduction
    • Sensorineural

    Strain Origin

    Donor
    Richard Cheney, Ph.D., UNC Chapel Hill School of Medicine.
    Primary Reference

    Heimsath EG Jr, Yim YI, Mustapha M, Hammer JA, Cheney RE. Myosin-X knockout is semi-lethal and demonstrates that myosin-X functions in neural tube closure, pigmentation, hyaloid vasculature regression, and filopodia formation. Sci Rep. 2017 Dec 11;7(1):17354. doi: 10.1038/s41598-017-17638-x. (Medline PMID: 29229982)

    Colony and Husbandry Information

    Special Considerations

    Because approximately half of homozygous Myo10tm1a embryos exhibit the lethal birth defect of exencephaly, it takes 2-3x more breeding effort to generate a given number of homozygous mice, although the homozygotes that survive birth appear to grow and breed normally. To maintain colonies and generate homozygotes, it can be helpful to mate homozygous males with heterozygous females, especially if embryo harvest is required.

    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
    On a B6 background, virtually all homozygous Myo10 KO mice, including Myo10tm1a, have one or more small white spots or flecks on the belly and/or back on an otherwise black coat; heterozygotes lack white spots and appear normal.
    Eye
    Black

    Breeding

    MMRRC Breeding System
    Random intra-strain mating
    Generation
    Not known precisely; minimuga testing in May 2021 revealed 99.8% consistent with C57BL/6J and 0% inconsistent.
    Overall Breeding Performance
    Good

    Reproductive Statistics

    Viability and Fertility: Female Male Comments
    Homozygotes are viable: Reduced Reduced
    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.

    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.

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

    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
    071696-UNC-RESUS Litter recovered from cryo-archive $3,088.00 / 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.