Description

Strain Information

Type Congenic; Mutant Strain; Targeted Mutation; Additional information on Genetically Engineered Mutant Mice. Species laboratory mouse Generation N11F1N1p (30-MAY-04)   Donating Investigator Peter Sorger,   Massachusetts Institute of Technology

Description
Mice that are heterozygous for the Mad2l1^tm1Sorg targeted allele are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. Homozygous null mice are embryonic lethal; they fail to develop past embryonic days 6-7 due to catastrophic chromosome missegregation and apoptosis. Histological analysis of heterozygous mutant mice reveal increased germinal center formation in spleen and a higher incidence of lung carcinomas when compared to the wildtype. This mutant mouse strain represents a model that may be useful in studies of the role cell-division checkpoints in tumorogenesis.

Development
A targeting vector containing neomycin resistance gene driven by the mouse phosphoglycerate kinase promoter and herpes simplex virus thymidine kinase genes was used to disrupt all coding sequence of the targeted gene. The construct was electroporated into 129S2/SvPas derived D3 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts.

Control Information

	Control
	Wild-type from the colony
	000664 C57BL/6J
Considerations for Choosing Controls

Additional Web Information

Congenic Nomenclature

Phenotype

Phenotype Information

View Mammalian Phenotype Terms

Mammalian Phenotype Terms

      assigned by genotype

The following phenotype information may relate to a genetic background differing from this JAX^® Mice strain.

Mad2l1^tm1Sorg/Mad2l1⁺

        involves: 129S2/SvPas * C57BL/6

• tumorigenesis
• lung adenocarcinoma (MGI Ref ID J:67456)
  • at 18-19 months of age, heterozygotes show a high frequency of papillary lung adenocarcinomas relative to age-matched wild-type mice
  • in contrast, the background rate of lymphomas detected in ageing heterozygotes remains unchanged relative to wild-type
• cellular phenotype
• *normal* cellular phenotype (MGI Ref ID J:62829)
  • initial studies indicated that heterozygotes develop normally, with blastocysts exhibiting no evidence of a defective mitotic checkpoint as assayed by reactivity to phospho-histone 3 antibody; premature sister-chromatid separation was not examined in initial experiments
• chromosome breakage (MGI Ref ID J:67456)
  • MEFs from 4 out of 5 heterozygotes exhibit high frequencies of premature anaphase (sister-chromatid separation) relative to wild-type cultures
  • all heterozygous MEF cultures display a significant increase in the number of aneuploid cells relative to wild-type cultures
  • the % of aneuploid cells correlates with the severity of premature anaphase, suggesting a direct relationship between the severity of loss of checkpoint control and chromosome mis-segregation
• hematopoietic system phenotype
• increased spleen germinal center number (MGI Ref ID J:62829)
  • heterozygotes are viable and largely normal but exhibit atypical (increased) germinal center formation in spleen relative to wild-type mice
• immune system phenotype
• increased spleen germinal center number (MGI Ref ID J:62829)
  • heterozygotes are viable and largely normal but exhibit atypical (increased) germinal center formation in spleen relative to wild-type mice

Mad2l1^tm1Sorg/Mad2l1⁺

        involves: 129S2/SvPas

• cellular phenotype
• abnormal cell physiology (MGI Ref ID J:117337)
  • 7.2% of MEFs are aneuploid for chromosome 2 compared to 1.4% of wild-type MEFs
  • mutant MEFs injected into athymic nude mice produced tumors in 8/8 mice within 4 weeks compared to no tumors from wild-type MEFs; tumors are 40% smaller at 6 weeks compared to than produced by MEFs from Mad1/Mad2 mutants
• abnormal chromosome number (MGI Ref ID J:117337)
  • chromosomal instability is much higher than in wild-type MEFs
• aneuploidy (MGI Ref ID J:117337)
  • at 5 months of age 15% of splenocytes are aneuploid; however no early aging related phenotypes are seen in heterozygotes at 19 to 27 months of age and no increase in the number of senescent MEFs is seen compared to wild-type
• tumorigenesis
• increased tumor incidence (MGI Ref ID J:117337)
  • mutant MEFs induce tumors in 100% of athymic nude mice in 4 weeks

Mad2l1^tm1Sorg/Mad2l1^tm1Sorg

        involves: 129S2/SvPas * C57BL/6

• lethality-prenatal/perinatal
• embryonic lethality before somite formation (MGI Ref ID J:62829)
  • homozygous mutant embryos die in utero between E6.5 and E7.5
• cellular phenotype
• abnormal cell cycle checkpoint function (MGI Ref ID J:62829)
  • at E5.5, mutant embryonic cells assemble normal spindles and undergo mitosis but are unable to arrest in response to drug-induced spindle disruption, indicating loss of a functional spindle assembly checkpoint
• abnormal mitosis (MGI Ref ID J:62829)
  • strikingly, ~25% of mitotic cells contain one or more lagging chromosomes, suggesting that anaphase proceeds in the absence of complete chromosome-microtubule attachment
  • in utero, E6.5-E7.5 mutant embryos show a significant reduction in the total number of mitotic cells
• chromosome breakage (MGI Ref ID J:62829)
  • absence of a functional spindle assembly checkpoint allows mutant cells to proceed even when unattached chromosomes are present, resulting in widespread chromosome missegregation
• increased apoptosis (MGI Ref ID J:62829)
  • mutant embryos undergo programmed cell death after the initiation of gastrulation at E6.5-E7.5; at this stage, wild-type embryos contain only a few apoptotic cells near the embryo center
  • in vitro, cell death is restricted to the rapidly dividing cells of the inner cell mass
• embryogenesis phenotype
• *normal* embryogenesis phenotype (MGI Ref ID J:62829)
  • homozygous mutant embryos appear normal both in utero and in culture until E5.5
• abnormal embryonic tissue morphology (MGI Ref ID J:62829)
  • mutant embryos appear grossly abnormal relative to wild-type embryos
• abnormal inner cell mass (MGI Ref ID J:62829)
  • in vitro, the ICM of mutant embryos stops proliferating after E6.5 and virtually no mutant ICM cells persist to E8.5
  • in contrast, the postmitotic and highly polyploid trophoblast giant cells continue to grow in size through E8.5
• reduced embryo size (MGI Ref ID J:62829)
  • mutant embryos are small relative to wild-type embryos
• growth/size phenotype
• reduced embryo size (MGI Ref ID J:62829)
  • mutant embryos are small relative to wild-type embryos

View Research Applications

Research Applications

This mouse can be used to support research in many areas including:

Mad2l1^tm1Sorg related

Apoptosis Research
Endogenous Regulators

Cancer Research
Increased Tumor Incidence (Other Tissues/Organs: lung)

Cell Biology Research
Cell Cycle Regulation

Developmental Biology Research
Embryonic Lethality (Homozygous)

Genes & Alleles

Gene & Allele Information

Allele Symbol		Mad2l1tm1Sorg
Allele Name		targeted mutation 1, Peter K Sorger
Allele Type		Targeted (knock-out)
Common Name(s)		Mad2-;
Mutation Made By		Max Dobles,   Biogen Inc
Strain of Origin		129S2/SvPas
ES Cell Line Name		D3
ES Cell Line Strain		129S2/SvPas
Gene Symbol and Name		Mad2l1, MAD2 (mitotic arrest deficient, homolog)-like 1 (yeast)
Chromosome		6
Gene Common Name(s)		AA673185; HSMAD2; MAD2; expressed sequence AA673185;
Molecular Note		A PGK-neomycin resistance cassette replaced the entire coding region. [MGI Ref ID J:62829]

Genotyping

Genotyping Information

Genotyping Protocols

Mad2l1^tm1Sorg, STD PCR, vers. 1

Helpful Links

Optimizing PCR Protocols

References

References

Selected Reference(s)

Dobles M; Liberal V; Scott ML; Benezra R; Sorger PK. 2000. Chromosome missegregation and apoptosis in mice lacking the mitotic checkpoint protein Mad2. Cell 101(6):635-45. [PubMed: 10892650]  [MGI Ref ID J:62829]

Additional References

Mad2l1^tm1Sorg related

Baker DJ; Jeganathan KB; Malureanu L; Perez-Terzic C; Terzic A; van Deursen JM. 2006. Early aging-associated phenotypes in Bub3/Rae1 haploinsufficient mice. J Cell Biol 172(4):529-40. [PubMed: 16476774]  [MGI Ref ID J:105717]

Burds AA; Lutum AS; Sorger PK. 2005. Generating chromosome instability through the simultaneous deletion of Mad2 and p53. Proc Natl Acad Sci U S A 102(32):11296-301. [PubMed: 16055552]  [MGI Ref ID J:100464]

Iwanaga Y; Chi YH; Miyazato A; Sheleg S; Haller K; Peloponese JM Jr; Li Y; Ward JM; Benezra R; Jeang KT. 2007. Heterozygous deletion of mitotic arrest-deficient protein 1 (MAD1) increases the incidence of tumors in mice. Cancer Res 67(1):160-6. [PubMed: 17210695]  [MGI Ref ID J:117337]

Michel LS; Liberal V; Chatterjee A; Kirchwegger R; Pasche B; Gerald W; Dobles M; Sorger PK; Murty VV; Benezra R. 2001. MAD2 haplo-insufficiency causes premature anaphase and chromosome instability in mammalian cells. Nature 409(6818):355-9. [PubMed: 11201745]  [MGI Ref ID J:67456]

Niault T; Hached K; Sotillo R; Sorger PK; Maro B; Benezra R; Wassmann K. 2007. Changing mad2 levels affects chromosome segregation and spindle assembly checkpoint control in female mouse meiosis I. PLoS ONE 2(11):e1165. [PubMed: 18043727]  [MGI Ref ID J:130342]

Health & husbandry

Health & Colony Maintenance Information

Colony Maintenance

Breeding & Husbandry

The resulting chimeric animals were crossed to C57BL/6 mice, and then backcrossed to the same for 10 generations (3/02). This strain is maintained as a heterozygote. Homozygous mutant mice have an embryonic lethal phenotype, failing to develop past embryonic days 6-7. Donating Investigator reports reproductive performance of heterozygotes is normal and recommends breeding heterozygotes at approximately 2 months of age; continue to breed for at least 18 months.

Diet Information

LabDiet® 5K52/5K67

Purchasing information

Pricing, Supply Level & Notes, Controls, General Terms & Conditions

Pricing

Supply Details

Standard Supply

Repository-Cryopreserved. Must Be Recovered. Please refer to pricing and supply notes for further information.

Supply Notes

Cryorecovery - Standard. The recovery process begins when a signed agreement form is returned to the Customer Service Department after order placement. Although results vary by strain, at least two males and two females (two pairs) will be provided, typically within 15 weeks of our receipt of the signed agreement form. If the first recovery attempt is unsuccessful or only one pair is recovered, a second recovery will be done, extending the delivery time to approximately 25 weeks. At least one member of each pair will be of known genotype and will carry the mutation if it is a mutant strain. Please note that pairs may not reflect the mating scheme utilized by The Jackson Laboratory prior to cryopreservation of the strain. Mating schemes are sometimes modified for successful cryopreservation. Price represents a repository maintenance fee, which includes the cost of recovery of the strain from the cryopreservation resource and the periodic replacement of the frozen embryos used for recovery. Cryorecovery to establish a Dedicated Supply for greater quantities of mice. One to two pairs will be recovered to establish a Dedicated Supply of mice. Price by quotation. For more information on Dedicated Supply, please contact JAX® Services, Tel: 1-800-422-6423 or 1-207-288-5845. This strain is included in the Induced Mutant Resource Colony collection.

Control Information

	Control
	Wild-type from the colony
	000664 C57BL/6J
Considerations for Choosing Controls
USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains.
International - Control Pricing Information for Genetically Engineered Mutant Strains.

General Terms and Conditions

See Terms of Use

The Jackson Laboratory's Genotype Promise

The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX^® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX^® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX^® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

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Ordering and Purchasing Information

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Contact Information
Orders & Technical Support
Tel: 800.422.6423 or 207.288.5845
Fax: 207.288.6150
Technical Support Email Form

Terms of Use

Terms of Use

General Terms and Conditions

For Licensing and Use Restrictions view the link(s) below:
- Use of MICE by companies or for-profit entities requires a license prior to shipping.

Contact information

General inquiries

Contracts Administration

phone:	207-288-6470
fax:	207-288-6655

JAX^® Mice & Services Conditions of Use

“Each recipient institution, including its employees and other researchers under its control (RECIPIENT), of mice or services using mice from The Jackson Laboratory (TJL) agrees that such mice, descendants of those mice derived by inbreeding or crossbreeding, including unmodified derivatives of those mice or their descendants (“MICE”) shall not be: (i) used for any purpose other than the internal research of the RECIPIENT, (ii) sold or otherwise provided to any third party for any use, or (iii) provided to any agent or other third party to provide breeding or other services with respect to MICE. Acceptance of MICE from TJL shall be deemed agreement by RECIPIENT to these conditions, and departure from these conditions requires The Jackson Laboratory’s prior written authorization.”

In case of dissatisfaction for a valid reason and claimed in writing by a purchaser within ninety (90) days of receipt of MICE, products or services, The Jackson Laboratory will, at its option, provide credit or replacement for the MICE or product received or the services provided.

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MICE and biological materials are to be used in a safe manner and in accordance with all applicable governmental rules and regulations.

The foregoing represents the General Terms and Conditions applicable to The Jackson Laboratory’s MICE, products and services. In addition, special terms and conditions of sale of certain MICE, products and services may be set forth separately in The Jackson Laboratory web pages, catalogs, price lists, contracts, and/or other documents, and these special terms and conditions shall also govern the sale of these MICE, products and services by The Jackson Laboratory, and by its licensees and distributors.

Acceptance of delivery of MICE, products or services shall be deemed agreement to these terms and conditions. No purchase order or other document transmitted by purchaser or recipient that may modify the terms and conditions hereof, shall be in any way binding on The Jackson Laboratory, and instead the terms and conditions set forth herein, including any special terms and conditions set forth separately, shall govern the sale of MICE, products services by The Jackson Laboratory.