CBA;B6-Tg(Sox10-icre/ERT2)388Wdr/J

Stock number: 027651
Product name: Sox10-iCreERT2
Research areas: Neurobiology Research, Research Tools

Description

These Sox10-iCreER^T2 mutant mice, carrying a transgene with tamoxifen-inducible optimized (improved) Cre recombinase under the control of the mouse Sox10 (SRY (sex determining region Y)-box 10) promoter, expresses iCreER^T2 in oligodendrocyte lineage cells.

Detailed description

The transcription factor encoded by the Sox10 gene is important in neural crest development, melanocyte development and homeostasis, myelination and mammary epithelial cell development. These Sox10-iCreER^T2 mutant mice carry a transgene with tamoxifen-inducible optimized (improved) Cre recombinase under the control of the mouse Sox10 (SRY (sex determining region Y)-box 10) promoter. iCre is an "improved Cre", optimized with mammalian codon usage, removed putative cryptic splice sites, an altered stop codon, and reduced CpG content to limit the chances of epigenetic silencing in mammals. When crossed with a strain containing a loxP site flanked sequence of interest, the offspring are useful for generating tamoxifen-induced, iCre-mediated targeted deletions. Tamoxifen administration induces Cre recombination in oligodendrocyte lineage cells. When crossed to a ROSA-YFP reporter strain, the resulting double mutant mice, treated with tamoxifen, exhibited YFP expression in more than 90% of Sox10 expressing cells in the spinal cord, and 65% of Sox10 expressing cells in the cerebral cortex and corpus callosum one week after treatment. Homozygotes are viable and fertile. The Donating Investigator reports that there is no difference in phenotype between homozygotes and hemizygotes.

The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17ß-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, Cre-ERT2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered.

Development

The 127 kb mouse phage artificial chromosome (PAC) RP21-427F18 containing the entire Sox10 gene, was modified by replacing the Sox10 open reading frame with an iCreERT2 fusion gene (an optimized variant of Cre recombinase fused to Cre-ER^T2). DNA from the modified PAC clone was used as the source for the transgene, which was microinjected into the pronucleus of CBA;B6 fertilized eggs. No other genes on the RP21-427F18 PAC were modified or deleted. Founder line 388 was subsequently established. 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).

Allele

Symbol: Tg(Sox10-icre/ERT2)388Wdr
Name: transgene insertion 388, William D Richardson
MGI accession ID: MGI:5634390
Generation method: Transgenic
Attributes: Recombinase-expressing; Inducible
Synonyms: Sox10-iCreER^T2; Tg(Sox10-icre/ERT2)#aWdr
Marker symbol: Tg(Sox10-icre/ERT2)388Wdr
Marker name: transgene insertion 388, William D Richardson
Chromosome: UN
Strain of origin: C57BL/6 or CBA
Expressed genes: cre/ERT2,Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of expression: iCreERT2 is expressed in oligodendrocyte lineage cells.
Molecular note: The transgene was generated using the PAC RP21-427F18. Oligodendrocyte-specific expression of the inducible improved cre recombinase (icre/ERT2) was achieved by replacing the coding sequence of Sox10 with the fusion. Four founders were obtained and one (388) was selected for further study.