STOCK Gt(ROSA)26Sor^{tm3(CAG-SOX9)Vlf}/J

Stock number: 039627
Product name: R26^SOX9
Research areas: Developmental Biology Research, Internal/Organ Research, Research Tools

Description

R26^SOX9 contain a Cre-inducible human SOX9 cDNA, with an N-terminal 3xFLAG tag, inserted into the Gt(ROSA)26Sor locus. These mice may be useful when studying the role of SOX9 in embryonic development and determination of the cell fate.

Detailed description

R26^SOX9/SOX9 mice contain a Cre-inducible knock-in of the human SRY (sex-determining region Y)-box 9 (SOX9) cDNA, with an N-terminal 3xFLAG tag, inserted into the Gt(ROSA)26Sor locus. A floxed Neo/STOP cassette prevents 3xFLAG-SOX9 expression in the absence of Cre recombinase. After removal of the loxP-flanked cassette, 3xFLAG-SOX9 is produced in Cre-expressing cells and their daughter cells.

Sox9 encodes a transcription factor involved in the regulation of lineage fate determination and differentiation of discrete cell types from embryonic development onto adulthood. SOX9 is highly expressed in the chondrocyte lineage and required for the survival and multi-step differentiation of the cells. It is thereby required for cartilage formation and maintenance. SOX9 is also highly expressed and has key roles in Sertoli cells, and is thereby involved in male differentiation. Additionally, SOX9 is expressed and has important roles in many types of progenitor, neural crest, glial and neuronal cells.

R26^SOX9/SOX9 mice are viable, look normal throughout life and are fertile.

R26^SOX9/+ mice mice carrying a Prx1Cre transgene (Stock No. 005584), which is active in early limb bud mesenchymal cells and some sternum progenitor cells, are viable, fertile and behave normally, but show a mild delay in endochondral ossification. R26^SOX9/SOX9Prx1Cre mice exhibit very short limbs with midshaft bone fractures due to cartilage growth plate defects and severely impaired bone formation. They show a drastic health decline in the second week postnatally, likely as a consequence of their multiple limb bone fractures.

Development

The R26^SOX9 targeting vector was designed with (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP-flanked stop cassette containing a neomycin resistance (neo) cassette and three polyadenylation signals, and 3xFLAG tagged human SRY (sex determining region Y)-box 9 (SOX9) cDNA sequence followed by a polyadenylation signal. This targeting vector also included 5' and 3' arms of homology for site-directed recombination into the Gt(ROSA)26Sor locus. The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1-derived C2 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and the chimeric mice were bred to C57BL/6J mice (Stock No. 000664) for at least 5 generations by the donating laboratory (see SNP note below). Cryopreserved sperm was sent to The Jackson Laboratory. Upon arrival, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes to establish our live colony.

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. Four of the 43 markers throughout the genome were segregating with an unknown source. Also, all 3 markers throughout the genome were segregating with 129. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed genetic background.

Allele

Symbol: Gt(ROSA)26Sor^{tm3(CAG-SOX9)Vlf}
Name: targeted mutation 3, Veronique Lefevbre
MGI accession ID: MGI:7665210
Generation method: Targeted
Attributes: Inserted expressed sequence; Epitope tag
Synonyms: R26^SOX9
Marker symbol: Gt(ROSA)26Sor
Marker name: gene trap ROSA 26, Philippe Soriano
Marker synonyms: beta geo; Gtrgeo26; Gtrosa26; R26; ROSA26; SETD5-AS1; Thumpd3as1
Chromosome: 6
Strain of origin: (C57BL/6 x 129S4/SvJae)F1
Molecular note: The targeting vector was designed to insert (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP site flanked stop cassette containing a neomycin resistance (neo) cassette and three polyadenylation signals, and 3xFLAG tagged human SRY (sex determining region Y)-box 9 (SOX9) cDNA sequence followed by a polyadenylation signal into the Gt(ROSA)26Sor locus.