B6;129-Llgl1^tm1Vv/J

Stock number: 020159
Product name: Llgl1 floxed
Research areas: Cell Biology Research, Developmental Biology Research, Neurobiology Research, Research Tools

Description

These floxed mutant mice possess loxP sites flanking exon 2 of the Llgl1 gene. This strain may be useful for generating conditional mutations in applications related to the study of cell polarity.

Detailed description

The targeted Llgl1 gene encodes an evolutionarily conserved protein involved in cell polarity maintenance, and cytoskeletal structure such as apical junction complexes. Mutations in this gene have been associated with Smith-Magenis Syndrome.

These mice possess loxP sites on either side of exon 2 of the targeted gene. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues. Removal of the floxed sequence creates a frameshift resulting in production of only the first 27 amino acids, and a null allele.

When bred to a strain with Cre recombinase expression in early embryos (see Stock No. 003755 for example), this mutant mouse strain may be useful in studies of embryonic development of the brain, loss of cell polarity and regulation of hematopoietic stem cells.

Development

A targeting vector containing a loxP site flanked SA-IRES beta-geo selection cassette was utilized in the construction of this mutant. This selection cassette was inserted upstream of exon 2 of the targeted gene, and another loxP site was inserted downstream of exon 2. Exon 2 is the exon downstream of the exon with the first ATG codon. This construct was electroporated into unspecified 129-derived embryonic stem (ES) cells which were transiently transfected with a Cre recombinase vector to remove the selection cassette. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were backcrossed to C57BL/6 for 8 generations by the donating lab (see SNP notes below). Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 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. Eleven of the 43 markers throughout the genome were segregating, suggesting an incomplete backcross.

Allele

Symbol: Llgl1^tm1Vv
Name: targeted mutation 1, Valeri Vasioukhin
MGI accession ID: MGI:3039268
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); No functional change
Marker symbol: Llgl1
Marker name: LLGL1 scribble cell polarity complex component
Chromosome: 11
Strain of origin: 129
Molecular note: The exon immediately downstream of the first coding exon was left flanked by single loxP sites following cre-mediated excision of an IRES-betageo cassette used for selection.