B6.129S6-Ep300^tm3Pkb/J

Stock number: 025169
Product name: p300 delta CH1
Research areas: Cell Biology Research, Developmental Biology Research, Metabolism Research

Description

Portions of the CH1 domain were deleted from the Ep300 gene to create these hypomorphic p300 deltaCH1 mice that are useful in studies of hypoxia-responsive transcription.

Detailed description

Ep300 (E1A binding protein p300) is a histone acetylase that is required for normal development. It interacts with a significant percentage of mammalian transcriptional regulatory proteins and has a CH1 domain that binds the C-terminal activation domain (C-TAD) of the hypoxia-inducible transcription factors HIF-1α and HIF-2α. This interaction is highly essential for hypoxia-responsive transcription, and is relevant to processes of glucose metabolism, angiogenesis, hematopoiesis, cell survival, invasion, and vascular tone.

These mice carry a targeted, partial, in-frame deletion of the CH1 domain in the mouse Ep300 gene (amino acids 329-379) that results in a hypomorphic protein. Western blots indicate that normal levels of a stable protein with intact HAT domain are produced. This strain was backcrossed to C57BL/6J for 25 generations by the donating lab.

Homozygotes on a random mixed background incorporating C57BL/6 and 129 alleles are overtly normal, although they are produced at about 50% the expected frequency. Two separate lines, one backcrossed to C57BL/6J (this strain), the other backcrossed to 129S2/SvEvTac (see Stock No. 025527), were generated by the donating laboratory. Homozygous mutants generated from an F1 cross of these two backgrounds are viable and have increased metabolic control. They are lean and demonstrate increased insulin sensitivity.

Homozygous viability and phenotype on the C57BL/6 background have not been assessed.

Development

Portions of the CH1 domain (amino acids 329-379) encoded by exon 4 were deleted and a neomycin resistance cassette flanked by FRT sites was introduced to the downstream intron through homologous recombination in 129S6/SvPasCrl-derived TL1 embryonic stem (ES) cells. Transient infection of the ES cells with a Flp-expressing vector excised the neomycin cassette. This strain was backcrossed to C57BL/6J for 25 generations by the donating laboratory.

Allele

Symbol: Ep300^tm3Pkb
Name: targeted mutation 3, Paul K Brindle
MGI accession ID: MGI:3612049
Generation method: Targeted
Marker symbol: Ep300
Marker name: E1A binding protein p300
Chromosome: 15
Strain of origin: 129S6/SvEvTac
Molecular note: Portions of the CH1 domain (amino acids 329-379) encoded by exon 4 were deleted and a neomycin resistance cassette flanked by FRT sites was introduced to the downstream intron. Transient infection of the ES cells with a Flp-expressing vector excised the neomycin cassette. RT-PCR confirmed the deletion in mutant MEFs.