129-Mcm2^tm2Scpr/J

Stock number: 017612
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research, Research Tools

Description

Mcm2^IRES-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.

Detailed description

Mcm2^IRES-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 Mcm2^IRES-EGFP mutant allele is normal (as determined by Northern blots and immunological tests).

Homozygous Mcm2^IRES-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 Mcm2^IRES-EGFP mice may be useful for visualizing and recovering proliferation competent cells.

Development

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.

Allele

Symbol: Mcm2^tm2Scpr
Name: targeted mutation 2, Steven C Pruitt
MGI accession ID: MGI:3820530
Generation method: Targeted
Attributes: Reporter
Marker symbol: Mcm2
Marker name: minichromosome maintenance complex component 2
Chromosome: 6
Strain of origin: 129S6/SvEvTac
Expressed genes: 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.
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.