Strain Name:
C.129S6(Cg)-Ndufa1tm1.1Nay/Mmjax
Stock Number:
043917-JAX
Citation ID:
RRID:MMRRC_043917-JAX
Other Names:
Ndufa1S55A-129Svj, Ndufa1S55A-Balb/c, Ndufa1 knock-in model for Complex I deficiency

Strain Information

Ndufa1tm1.1Nay
Name: NADH:ubiquinone oxidoreductase subunit A1; targeted mutation 1.1, Nagendra Yadava
Synonyms: Ndufa1S55A
Type: Allele
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: Knock-In
Ndufa1
Name: NADH:ubiquinone oxidoreductase subunit A1
Synonyms: 1810049F12Rik, MWFE
Type: Gene
Species: Mus musculus (mouse)
Chromosome: X
Alteration at locus: Knock-In
NCBI: 54405
HGNC: HGNC:7683
Homologene: 3337
Genetic Alterations
Missense mutation in the Ndufa1 gene (MGI:1929511) in codon 55 (T>G; codon TCT>GCT), replacing serine-55 with alanine (p.S55A) in the MWFE subunit of Complex I.

Alteration details:
Genotype Determination
ES Cell Line
iTL1 derived from 129S6/SvEvTac
Phenotype

Homozygous: Mutant females show improved survival despite the presence of systemic partial respiratory chain Complex I deficiency (published). Abnormal mammary gland ductal architecture (unpublished)

Hetero/Hemizygous: Mutant males display glucose intolerance (unpublished) and reduced respiratory exchange ratio (RER) and produced less body heat. They are hypo-active and eat less. Despite eating less their weight gain is comparable or relatively more. The showed age-dependent Purkinje neurons' degeneration.

MeSH Terms
  • Animals
  • Body Temperature/physiology
  • Electron Transport Complex I/deficiency
  • Electron Transport Complex I/genetics
  • Exhalation/physiology
  • Female
  • Genes, X-Linked/genetics
  • Genetic Predisposition to Disease/genetics
  • Male
  • Membrane Proteins/genetics
  • Mice
  • Mice, 129 Strain
  • Mice, Transgenic
  • Neurodegenerative Diseases/genetics
  • Neurodegenerative Diseases/metabolism
  • Pregnancy
Strain Development
129/SvEv-derived male embryonic stem cells (ES) were used to target the Ndufa1S55A allele on its native locus at X-chromosome. The Ndufa1S55A allele harbors a point mutation in the codon-55 (T>G; codon TCT>GCT). The long homology arm was 6.0 kb upstream to the point mutation (T>G) in exon 2. The LoxP/FRT-Neo cassette for G418 selection was inserted 903 bp downstream of the point mutation within intron 2-3. The short homology arm extended 2.0 kb from the 5’-end of the Neo cassette. The T>G mutation was created by site-directed mutagenesis. ES clones with correct targeting by homologous recombination were screened first by polymerase chain reaction (PCR) and later confirmed by Southern blot analysis with an internal probe. Genomic DNA was digested with HpaI, and electrophoretically separated on a 0.8% agarose gel. After transfer to a nylon membrane, the digested DNA was hybridized with a probe targeted against the 3’ internal region. DNA from 129/SvEv was used as wild-type control. The expected sizes for wild-type (WT; 9.3 kb) and knock-in (KI Neo: 11 kb) were observed. Because ES cells were derived from male mice, only one copy of the Ndufa1 gene was expected. Therefore, correct targeting will result in only one band of 11 kb. By Southern blot analysis clone #253 was identified to have Ndufa1S55A allele correctly targeted at its native locus. After confirming the reduced MWFE and Complex I levels in ES clone #253, it was injected into C57BL/6 blastocysts.

Chimeras with high percentage agouti coat color were mated to 129 FLP mice (JAX #003946) to remove the Neo-cassette and simultaneously generate the germ-line founder mice. A mutant male (Ndufa1S55A/Y) and two heterozygous females (Ndufa1S55A/+) were obtained. They were crossed with wild-type mice (129S1/SvImJ, JAX #02448) to expand the colony. The deletion of Neo-cassette was confirmed by PCR screening using primers set F1 (5’-CAA AGC CAT TTC TCT GCC TCT-3’) and R1 (5’-CCA AAA AAC CTC ACT CAC AAC C-3’). The presence of Ndufa1S55A allele was confirmed using primers set F2 (5’ -TGA GCT ATC TCT TCA GCC TCT AG- 3’) and R2 (5’-GAC CCT GTC TCA ATT ACA AAT CAA CCA ACC -3’) by Hyp188III restriction fragment length polymorphism. The enzyme cuts only the wild-type (Ndufa1+) allele. Finally, the presence of T>G mutation in the Ndufa1S55A mice was confirmed by DNA sequencing. From gene targeting to founder mice generation, this project was supported by inGenious Targeting Laboratory (Stony Brook, NY) on a pay-for-service basis.

To transfer the Ndufa1S55A allele on BALB/c genetic background, we performed 9 generations of backcrossing. Because the Ndufa1 gene is X-chromosome linked, we crossed Ndufa1S55A-129S1 mutant males (r/Y) mice (from N5) with wild-type BALB/c females (X/X). Such a cross resulted in heterozygous females (r/X), which were mated with wild-type BALB/c males (X/Y). Then in the next generation, mutant male progeny (r/Y) were mated with wild-type BALB/c females (X/X). This alternating breeding scheme continued for another 8 generations (i.e., N9) to generate Ndufa1S55A-BALB/c strain.
Suggested Control Mice
Wild-type littermates
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 csmmrrc@jax.org. Older strains may not have this information.
  • Apoptosis
  • Cancer
  • Cardiovascular
  • Cell Biology
  • Developmental Biology
  • Diabetes
  • Endocrine Deficiency
  • Hematology
  • Immunology and Inflammation
  • Internal_Organ
  • Metabolism
  • Models for Human Disease
  • Neurobiology
  • Obesity
  • Research Tools
  • Sensorineural
Donor
Nagendra Yadava, Ph.D., Baystate Medical Center.
Primary Reference

Kim C, Potluri P, Khalil A, Gaut D, McManus M, Compton S, Wallace DC, YadavaN. An X-chromosome linked mouse model (Ndufa1(S55A)) for systemic partial ComplexI deficiency for studying predisposition to neurodegeneration and other diseases.Neurochem Int. 2017 Oct;109:78-93. doi: 10.1016/j.neuint.2017.05.003. Epub 2017May 12. (Medline PMID: 28506826)

Colony and Husbandry Information

Cryo-recovered strains distributed by the MMRRC at JAX are shipped to the customer from the Pathogen & Opportunistic-Free Animal Room G200 - see https://www.jax.org/jax-mice-and-services/customer-support/customer-service/animal-health/health-status-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 csmmrrc@jax.org.
Coat Color
agouti (A/A)
MMRRC Breeding System
Backcross
Breeding Scheme(s)
All possible breeding strategy can be used. Wild-type and mutant colony can be bred separately. In this case, no genotyping of progeny will be required. Alternatively, heterozygous females with wild-type males can be bred. However, in this case genotyping will be required.
Generation
N9 (BALB/c)
Overall Breeding Performance
Good
NOTE: "Hemizygote" as used here refers to males carrying a mutation on the X Chromosome or mice of either sex carrying an inserted transgene with no homologous allele on the other chromosome.
Viability and Fertility: Female Male Comments
Homozygotes are viable: Yes X-linked
Homozygotes are fertile: Yes Yes
Hetero/Hemizygotes are fertile: Yes Yes
Age Reproductive Decline: 4 to 6 months 4 to 6 months
Bred to Homozygosity
No
Average litter size
4-6
Recommended wean age
3 weeks
Average Pups Weaned
4-6

Order Request Information

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 csmmrrc@jax.org for more information.

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.

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
043917-JAX-SPERM Cryo-preserved spermatozoa $437.00 / $437.00
Non-Profit / For-Profit
Aliquot Approximate quantity3
043917-JAX-RESUS Litter recovered from cryo-archive $2,022.00 / $2,022.00
Non-Profit / For-Profit
Litter Recovered litter4; additional fees for any special requests.
Cryopreserved material may be available upon request, please inquire to csmmrrc@jax.org for more information.

Some MMRRC strains were submitted by the donor as cryopreserved germplasm, and because these strains were not cryopreserved by the MMRRC, we have not assessed the quality, nor genotype, of this material. Our expertise in reviving mice from various qualities of frozen sperm and embryos, using methods like IVF and ICSI, gives us confidence in successful recoveries in most cases. However, due to the uncertain quality of these samples, we'll limit revival attempts to two per order. Additional attempts are available for a fee, on top of standard charges, if requested. It's important to note that some strains may lack the expected mutation, so we can't assure successful order fulfillment until we attempt to revive the strain.

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.