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Strain Detail Sheet

Strain Name:
B6J.B6N(FVB)-Myo10tm1d(KOMP)Wtsi/Mmnc
Stock Number:
071698-UNC
Citation ID:
RRID:MMRRC_071698-UNC
Other Names:
Myo10tm1d

Strain Information

Gene Details

Myo10tm1d(KOMP)Wtsi
Name: myosin X; targeted mutation 1d, Wellcome Trust Sanger Institute
Synonyms: Myo10tm1d
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 15
Alteration at locus: Knockout
Myo10
Name: myosin X
Synonyms: myosin-X, D15Ertd600e
Type: Gene
Species: Mouse
Chromosome: 15
Alteration at locus: Knockout
NCBI: 17909
HGNC: HGNC:7593
Homologene: 36328

Additional Resources
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Genetic Alterations
The L1L2_Bact_P cassette was inserted at position 25785401 of Chromosome 15 upstream of the critical exon (exon 27) (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 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 exon 27 at position 25786400. Exon 27 is thus flanked by loxP sites. A null/knockout allele was created by flp and cre recombinase expression in mice carrying this allele to remove the lacZ sequence, neo selection cassette and the loxP-flanked exon 27. Immunoblots of whole brain from P5 pups confirmed that expression of both the full-length and headless forms is undetectable in homozygous mutant mice.
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 homozygous Myo10tm1d embryos exhibit the lethal neural tube defect of exencephaly. The tm1a homozygotes that are not affected by exencephaly exhibit nearly 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 adult tm1d homozygotes exhibit syndactyly. Based on the characterization reported in our manuscript on Myo10 null mice, the tm1a, tm1d, and m1j alleles exhibit a similar core phenotype, and the tm2 mice characterized by the IMPC exhibit a similar phenotype. It should be noted that the tm1a, tm1d, and m1j alleles delete full-length and headless Myo10 while the tm2 allele with its ~9 kb deletion spares at least one form of headless Myo10. 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 and some mice also have a white spot on their back. 100% of homozygotes whose eyes were dissected exhibited persistent fetal vasculature. Approximately 50% of Myo10tm1d homozygotes exhibited webbed digits. Hetero/Hemizygous: Heterozygotes appear normal and are not obviously different from wild-type. KO mice for experiments can be produced by mating homozygous males with heterozygous females.

    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
    KOMP Myo10tm1a "KO-first" ES cells (EPD0332_3-B02 cells derived from JMA3.N1 ES cells, a cell line derived from a male C57BL/6N-A tm1 Brd mouse; see KOMP project 50217) were injected into blastocysts. The resulting chimeras were bred to albino C57BL/6 mice to get germline transmission on a C57BL/6 background, producing a Myo10tm1a allele. The resulting germline were crossed with mice expressing FLPo recombinase to remove the "KO-first" cassette while retaining the floxed exon 27 to produce the Myo10tm1c allele. These mice were crossed with one another and with C57BL/6J mice prior to sending them to Cheney lab (UNC), where a colony was established by breeding them to one another prior to backcrossing to make the strain congenic to C57BL/6J (Jackson Strain #000664). MiniMuga testing in February of 2022 showed 99% C57BL/6J, so the mice were backcrossed more and then maintained by intra-strain mating of homozygotes.
    Suggested Control Mice
    WT littermates, C57BL/6J. Approximately half of homozygous Myo10tm1d embryos exhibit the lethal birth defect of exencephaly, it can be advantageous to increase the number of KO's obtained per litter by mating homozygous males to heterozygous females and using heterozygotes for 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 Myo10 KO mice exhibit the lethal embryonic defect of exencephaly, it typically takes 2-3x more breeding than normal to obtain a given number of homozygous Myo10tm1d mice, although the homozygous KO mice that survive birth appear to grow and breed normally. To maintain Myo10tm1dcolonies and generate homozygotes, it can be helpful to mate homozygous males to 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 Myo10tm1d, 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 certain
    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
    071698-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.