BAF250af/f floxed mice have loxP sites flanking exon 8 of the AT rich interactive domain 1A (SWI-like) (Arid1a) gene. This strain may be useful for generating conditional mutations for use in applications related to BAF250a-mediated chromatin remodeling and ES cell self-renewal, differentiation, and cell lineage decisions.
Zhong Wang, University of Michigan
BAF250af/f floxed mice possess loxP sites flanking exon 8 of the AT rich interactive domain 1A (SWI-like) (Arid1a) gene. BAF250a is a subunit of an ATP-dependent BAF chromatin remodeling complex, in the SNF/SWI superfamily. BAF250a is required for transcriptional activation of genes normally repressed by chromatin. Homozygotes are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 8 deleted in cre-expressing tissues. Studies of BAF250a-conditional KO embryos results in embryonic lethality showing the requirement for BAF250a during embryonic development.
A targeting vector was designed to insert a single loxP site upstream of exon 8, and a frt-flanked βgeo cassette, followed by a loxP site, downstream of exon 8 of the AT rich interactive domain 1A (SWI-like) (Arid1a) gene. The construct was electroporated into 129P2/OlaHsd-derived E14 embryonic stem (ES) cells. Correctly targeted ES cells were transiently transfected with a pCAGGsFLPe expression plasmid to delete the β-geo cassette. Resulting ES cells were injected into blastocysts and resulting chimeric BAF250af/f mice were intercrossed by the donating lab (see SNP note below). Upon arrival, mice were bred to 129S1/SvImJ inbred mice (Stock No. 002448) for at least one generation to establish the colony.
A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. Nine of the 27 markers throughout the genome were segregating, suggesting an unknown genetic contribution on multiple chromosomes.
|Allele Name||targeted mutation 1.1, Zhong Wang|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Arid1a, AT rich interactive domain 1A (SWI-like)|
|Strain of Origin||129P2/OlaHsd|
|Molecular Note||An frt-flanked betageo cassette with a 3' loxP site and a 5' SV40-polyA tail was inserted downstream of exon 8 and an additional loxP site was inserted upstream of exon 8. Transient flp expression in ES cells was used to remove the betageo cassette and leave exon 8 floxed.|
When maintaining a live colony mice homozygous for the floxed allele may be bred together.
When using the BAF250af mouse strain in a publication, please cite the originating article(s) and include JAX stock #027717 in your Materials and Methods section.