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Strain Name:
B6J;B6N-Mfrptm1.1Rayy/Mmucd
Stock Number:
075987-UCD
Citation ID:
RRID:MMRRC_075987-UCD
Other Names:
Mfrp KI/KI

Strain Information

QUALITY CONTROL OF B6J;B6N-Mfrptm1.1Rayy/Mmucd  |  University of California at Davis
Last Updated:
Strain Data and Information Information by Submitter Assessed by MMRRC1,2
Published Provided
Allele-specific genotype3n.d.
Genetic backgroundn.d.
Viability of genotypes available for distributionn.d.
Specific Pathogen- Status n.d.
Recoverability of cryopreserved sperm/embryos4 n.d.
Gene or allele sequence3 n.d.
Gene or allele expression3n.d.
Gene or allele function3n.d.
Observable and/or measurable phenotypesn.d.
Fertility of genotypes available for distributionn.d.
Fecundity/breeding performance n.d.

1 When indicated as verified ("YES"), then please note that information presented is to the best of our knowledge correct and up-to-date at the time of verification at the MMRRC Distribution Center; however, this information is subject to change due to breeding, maintenance, and other actions on the mouse strain at the MMRRC Distribution Center; direct any questions on this table to the MMRRC Distribution Center for this mouse stain.

2 If verification has not been performed (as indicated by "NO"), investigators may request specific verification testing for a fee. Requests should be submitted directly to the MMRRC Distribution Center assigned to the management, archiving, and distribution of the strain. A full listing of available testing and analytical services is available at https://www.mmrrc.org/about/services.php.

3 This information may or may not apply to each individual engineered allele (e.g., Cre, FlpO) present in the strain.

4 Recovery refers to thawing, in vitro fertilization (IVF), and/or embryo culture leading to live offspring.

Mfrptm1.1Rayy
Name: membrane frizzled-related protein; targeted mutation 1.1, Radha Ayyagari
Type: Allele
Species: Mus musculus (mouse)
Chromosome: 9
Alteration at locus: Knock-In
Mfrp
Name: membrane frizzled-related protein
Type: Gene
Species: Mouse
Chromosome: 9
NCBI: 259172
Homologene: 12866
Genetic Alterations
A vector containing exons 1-2, a loxP site, exons 3-9 with a 1 bp insertion of cytosine at c.498_499 (c.498_499insC) in exon 5, an FRT site, loxP site, neomycin cassette, FRT site, and loxP site, and exons 10-12 replaced exons 1-12. The neomycin cassette was removed via flp-mediated recombination, leaving exons 3-9 with the mutation in exon 5 floxed.

The homozygous c.498_499insC mutation has been identified in a family with hyperopia, microphthalmia, retinitis pigmentosa, retinal pigment epithelial atrophy, variable degrees of foveal edema, and opti disc drusen, newly described as MFRP-associated retinopathy. Mice free of the Crb1rd8 mutation were used to generate the colony.
ES Cell Line
iTL IC1 (ES Cell) derived from C57BL/6NTac
Phenotype

The model shares many of the features of human clinical disease, including reduced axial length, hyperopia, retinal degeneration, retinal pigment epithelial atrophy, and decreased electrophysiological responses. In addition, the eyes of Mfrp homozygous mice had a significantly greater refractive error (p < 0.01) when compared to age-matched wild-type control animals.

Administration of recombinant adeno-associated virus–mediated Mfrp gene therapy significantly prevented thinning from retinal neurodegeneration (p < 0.005) and preserved retinal electrophysiology (p < 0.001) when treated eyes were compared to contralateral sham-treated control eyes.

Conditional phenotype: Mfrp KI/KI mice carry loxp site to generate Mfrp knockout mice.

MeSH Terms
  • Animals
  • Biomarkers
  • Dependovirus/genetics
  • Disease Models, Animal
  • Electroretinography
  • Genetic Predisposition to Disease
  • Genetic Therapy/methods
  • Genetic Vectors/administration & dosage
  • Genetic Vectors/genetics
  • Humans
  • Immunohistochemistry
  • Membrane Proteins/genetics
  • Mice
  • Mice, Knockout
  • Phenotype
  • Retinal Diseases/diagnosis
  • Retinal Diseases/genetics
  • Retinal Pigment Epithelium/metabolism
  • Tomography, Optical Coherence
Strain GQC Summary
Gene Specific Genotyping:

To request gene-specific and other genotyping services for a strain, please contact the distribution MMRRC Center for more information.

Background Genetic Quality:

The MMRRC has developed a Genetic Quality Control pipeline using the MiniMUGA array to provide additional information to identify and validate genetic backgrounds of MMRRC strains. For more information on whether genetic background data is available, please contact MMRRC_GeneticQC@med.unc.edu. Note: that MiniMUGA genetic background data is not available on all strains, but can be ordered if desired.

Suggested Control Mice
Littermates of all relevant genotypes.
  • Neurobiology
  • Sensorineural
Submitter
Radha Ayyagari, Ph.D., University of California, San Diego.
Primary Reference

Chekuri A, Sahu B, Chavali VRM, Voronchikhina M, Soto-Hermida A, Suk JJ, Alapati AN, Bartsch DU, Ayala-Ramirez R, Zenteno JC, Dinculescu A, Jablonski MM, Borooah S, Ayyagari R. Long-Term Effects of Gene Therapy in a Novel Mouse Model of Human MFRP-Associated Retinopathy. Hum Gene Ther. 2019 May;30(5):632-650. doi: 10.1089/hum.2018.192. Epub 2019 Jan 16. (Medline PMID: 30499344)

Strain Development
The targeting vector was constructed by subcloning an 11.15 kb region from a C57BL/6 BAC clone (RP23:270P20). This 11.15 kb region included exons 1–13 of the Mfrp gene. A single LoxP site was inserted upstream of exon 3 in introns 2–3 and a LoxP/FRT-flanked Neo cassette was inserted downstream of exon 9 in introns 9–10.The C-insertion mutation in exon 5 was generated by site-directed mutagenesis. The target region was 2.7 kb containing exons 3–9 of the Mfrp gene. The total size of the targeting construct was 15.25 kb.

The targeting vector was linearized and transfected into iTL IC1 embryonic stem (ES) cells derived from C57BL/6NTac. Recombinant ES clones were screened by PCR following G418 antibiotic selection. The positive clone identified by PCR was further confirmed by Southern blotting analysis. The positive ES cell clones were injected into BALB/c blastocysts and implanted into pseudopregnant mice to generate chimeras. C57BL/6J mice were mated with these chimeras to transmit the targeting allele. Mfrp KI/KI mice were identified by PCR of tail-tip DNA.

In homozygous Mfrp KI mice, the kanamycin marker was flanked by an FRT sequence, and this was removed by breeding with FLP mice. Mice were tested for the rd8 mutation. Mice free of the rd8 mutation were utilized to generate the mouse colony with the homozygous c498_499insC mutation (Mfrp KI/KI).

The presence of a 1 bp insertion (cytosine) in Mfrp was confirmed by using a loxP specific forward and an endogenous Mfrp reverse primer.


Disclaimer: If MMRRC Strain Genetic Quality Control (GQC; based on MiniMUGA genotyping and analysis) has been completed for this strain, the information might differ from the genetic background information provided by the submitter. MiniMUGA genetic analysis is done on a strain’s tissue samples taken when archived by or ordered from the assigned MMRRC Center.

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@ucdavis.edu.
Coat Color
Black
Eye
Black
MMRRC Breeding System
Sib-mating
Generation
N/A
Overall Breeding Performance
Undetermined
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes Yes
Homozygotes are fertile: Yes Yes
Heterozygotes are fertile: Yes Yes
Age Reproductive Decline: Greater than 12 months Greater than 12 months
Average litter size
4 to 6
Recommended wean age
3 Weeks
Average Pups Weaned
4 to 6

Order Information

Limited quantities of breeder mice (recovered litter) are available from a cryoarchive; recovered litter usually available to ship in 3 to 4 months.

A Commercial License Agreement from the Submitter is required for for-profit entities to use this strain. For more information, please contact Eric Gosink.

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 Submitter is required for for-profit entities to use this strain. For more information, please contact Eric Gosink

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
075987-UCD-RESUS Litter recovered from cryo-archive $5,892.91 / $6,181.91
Non-Profit / For-Profit
Litter Recovered litter4; additional fees for any special requests.

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.