B6;129S4-E2f4^tm2.1Lees/J

Stock number: 031216
Product name: E2f4 f
Research areas: Research Tools

Description

Exons 2-4 of the mouse E2f4 gene are flanked by loxP sites. Cre-mediated excision of the floxed region results in a knock-out allele. These mice may be useful in studying cell cycle, cancer, organ development and homeostasis, and ciliopathies.

Detailed description

Dysfunction of primary cilia or multicilia results in human ciliopathies, conditions associated with high morbidity and mortality. Evidence suggests that multiciliogenesis requires E2f4/E2f5 (E2F transcription factor 4 and 5) activity - specifically for centriole biogenesis in multiciliated cell precursors. E2F transcription factors have also been associated with the regulation of entry/exit from cell cycle, and organ development/homeostasis. E2f4 expression has been identified as a prognostic marker in several cancers.

Exons 2-4 of the mouse E2f4 gene are flanked by loxP sites in this conditional mutant strain. Cre-mediated excision of the floxed region results in a knock-out allele.

Crosses with Shh-cre mice (see Stock No. 005622), expressing early in the lung epithelium, demonstrate that primary cilia formation is not prevented, but multiciliated cells fail to develop upon knock-out of E2f4.

When crossed with Vil-cre mice, expressing cre recombinase throughout the intestinal epithelium and efferent ducts of the testis from embryonic day 12.5, viability and fertility are normal. However, when E2f4 knock-out is combined with loss of one allele of E2f5 this leads to infertility and the loss of multiciliated cells from the efferent ducts. E2f4 is not required for normal intestinal development and homeostasis as the intestine appears morphologically normal.

Development

A FRT-flanked PGK-EM7-neo-bpA selection cassette containing one loxP site was introduced to intron 1 of the mouse E2f4 gene, and a second loxP site was placed in intron 4 via homologous recombination in (C57BL/6 x 129S4/SvJae)F1-derived v6.5 embryonic stem (ES) cells. Resultant chimeric male mice were crossed with C57BL/6 females. Subsequent crosses with mice expressing FLPe recombinase (see Stock No. 003946) excised the neo selection cassette, leaving exons 2-4 flanked by loxP sites. This strain was maintained on a mixed C57BL/6-129 genetic background by the donating laboratory.

Allele

Symbol: E2f4^tm2.1Lees
Name: targeted mutation 2.1, Jacqueline A Lees
MGI accession ID: MGI:5706731
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: E2f4
Marker name: E2F transcription factor 4
Chromosome: 8
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Molecular note: LoxP sites were introduced into intron 1 and intron 4. The positive selection cassette used for targeting was removed via FLPe mediated recombination after germline transmission.