B6;129S4-Sox9^tm1.1Tlu/J

Stock number: 030137
Product name: Sox9-IRES-EGFP KI
Research areas: Developmental Biology Research, Research Tools

Description

This Sox9^IRES-EGFP (or Sox9^IE) knock-in strain expresses EGFP from the endogenous Sox9 locus and is useful in studies of chondrogenesis and skeletal development.

Detailed description

The targeted Sox9 gene encodes a transcription factor important in chondrogenesis and skeletal development as well as sex determination. Mutations in this gene have been associated with Campomelic Dysplasia, Pierre Robin sequence, Cooks syndrome, and 46,XX sex reversal. These Sox9^IRES-EGFP (or Sox9^IE) mice carry a knock-in mutation in which IRES-EGFP has been inserted into the 3’UTR of the Sox9 locus. EGFP fluorescence is visible in homozygous E12.5 embryos in the expected Sox9 gene expression pattern. Sox9 and EGFP proteins are detected in homozygous E12.5 embryos by western blot analysis using anti-Sox9 and anti-GFP antibodies respectively. No Sox9-EGFP fusion protein is detected in western blots of homozygous E12.5 embryos. The EGFP in the knock-in allele allows isolation of Sox9 expressing cells using FACS. Mice that are homozygous for the targeted mutation are viable and fertile.

Development

A targeting vector was used to insert an IRES-EGFP-FRT-Neo-FRT into the 3’ UTR of the targeted gene. The construct was electroporated into (C57BL/6J x 129S4/SvJae)F1 derived v6.4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into 2-8 cell stage embryos generated from matings of CD1 wild type mice. The resulting chimeric animals were tested for germline transmission. The mice were crossed to Flpe recombinase expressing mice on the 129S4/SvJaeSor genetic background (Stock No. 003946) to excise the FRT site flanked neo cassette. The mice were then intercrossed to generate homozygotes. The mice no longer carry the neo selection cassette or the Flpe recombinase allele. 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: Sox9^tm1.1Tlu
Name: targeted mutation 1.1 Thomas Lufkin
MGI accession ID: MGI:5300542
Generation method: Targeted
Attributes: Reporter
Synonyms: Sox9^IE
Marker symbol: Sox9
Marker name: SRY (sex determining region Y)-box 9
Marker synonyms: 2010306G03Rik; CMD1; CMPD1; ENH13; SRA1; SRXX2; SRXY10; SRY; TESCO
Chromosome: 11
Strain of origin: (C57BL/6J x 129S4/SvJae)F1
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: EGFP is observed in Sox9 expression domains such as the vertebral column and limbs.
Molecular note: An IRES-EGFP-FRT-Neo-FRT is inserted into the 3' UTR. Flp-mediated recombination removed the neo cassette.