Mcm2IRES-EGFP mice have an enhanced green fluorescent protein (EGFP) sequence inserted downstream of the stop codon of the minichromosome maintenance deficient 2 mitotin (Mcm2) gene. These mice may be useful for visualizing and recovering proliferation competent cells.
Steven Pruitt, Roswell Park Cancer Institute
Genetic Background | Generation |
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Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Mcm2 | minichromosome maintenance complex component 2 |
Mcm2IRES-EGFP mice have an enhanced green fluorescent protein (EGFP) sequence inserted downstream of the stop codon of the minichromosome maintenance deficient 2 mitotin (Mcm2) gene. Mcm2 expression from the Mcm2IRES-EGFP mutant allele is normal (as determined by Northern blots and immunological tests).
Homozygous Mcm2IRES-EGFP mice are viable, fertile, normal in size, and do not display any gross physical or behavioral abnormalities. EGFP expression is seen in regions known to be highly proliferative; including spleen, small intestine, colon, and hair follicles. EGFP expression is also detectable in the majority of bone marrow and in slowly-cycling neural stem cells in the subventricular zone of the brain.
Mcm2 is a component of the DNA replication licensing complex expressed in stem/progenitor cells in a variety of regenerative tissues. Mcm2 plays an essential role in eukaryotic cell division as it marks DNA replication origins during G1 for use in the subsequent S-phase. MCM2 prevents multiple rounds of replication from a single origin and suppresses endoreduplication. Mice harboring other mutations resulting in reduced Mcm2 expression exhibit highly penetrant early onset cancers and stem cell deficiencies. These Mcm2IRES-EGFP mice may be useful for visualizing and recovering proliferation competent cells.
A targeting vector was designed to insert an internal ribosome entry site (IRES), and an enhanced green fluorescent protein (EGFP) sequence followed by a polyA signal and a neomycin resistance cassette downstream of the stop codon of the minichromosome maintenance deficient 2 mitotin (Mcm2) gene. The construct was electroporated into 129S6/SvEvTac-derived W4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to 129/Sv females. These mutant mice were maintained on a 129/Sv background. Upon arrival, mice were bred to 129S1/SvImJ mice (Stock No. 002448) for at least one generation to establish the colony.
Expressed Gene | GFP, Green Fluorescent Protein, |
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Site of Expression | EGFP expression is seen in regions known to be highly proliferative; including spleen, small intestine, colon, and hair follicles. EGFP expression is also detectable in the majority of bone marrow and in slowly-cycling neural stem cells in the subventricular zone of the brain. |
Allele Name | targeted mutation 2, Steven C Pruitt |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | Mcm2IRES-EGFP |
Gene Symbol and Name | Mcm2, minichromosome maintenance complex component 2 |
Gene Synonym(s) | |
Expressed Gene | GFP, Green Fluorescent Protein, |
Site of Expression | EGFP expression is seen in regions known to be highly proliferative; including spleen, small intestine, colon, and hair follicles. EGFP expression is also detectable in the majority of bone marrow and in slowly-cycling neural stem cells in the subventricular zone of the brain. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 6 |
Molecular Note | An IRES-EGFP and PGK-neo cassette was inserted into the XbaI restriction site found in exon 16 after the stop codon. Gene expression is normal in Northern blots and by immunological tests. |
Mutations Made By | Steven Pruitt, Roswell Park Cancer Institute |
When maintaining a live colony, homozygous mice may be bred together.
When using the 129-Mcm2tm2Scpr/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #017612 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Mcm2<tm2Scpr> |
Frozen Mouse Embryo | 129-Mcm2<tm2Scpr>/J | $2595.00 |
Frozen Mouse Embryo | 129-Mcm2<tm2Scpr>/J | $2595.00 |
Frozen Mouse Embryo | 129-Mcm2<tm2Scpr>/J | $3373.50 |
Frozen Mouse Embryo | 129-Mcm2<tm2Scpr>/J | $3373.50 |
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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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