B6.Cg-Tg(Col2a1-d2EGFP)1Waho/J

Stock number: 030672
Product name: Col2-pd2EGFP
Research areas: Research Tools

Description

A destabilized form of EGFP (d2EGFP) is expressed from the mouse Col2a1 promoter, providing a near real-time fluorescent tool for in vivo/in vitro studies of chondrocytes.

 

Detailed description

These transgenic mice express a destabilized d2EGFP fluorescent reporter from the mouse Col2a1 (collagen, type II, alpha 1) promoter/enhancer. d2EGFP, derived from plasmid pd2EGFP-N1, encodes a destabilized, red-shifted variant of wildtype GFP which has been optimized for brighter fluorescence and higher expression in mammalian cells. A half-life of 1-2 hours is reported, enabling near real-time in vivo/in vitro studies of chondrocytes.

In the post-natal growth plate, d2EGFP fluorescence is expressed in almost all proliferative chondrocytes and in some hypertrophic chondrocytes. In articular cartilage, d2EGFP fluorescence diminishes over time, demonstrating the decrease of type II collagen synthesis in articular chondrocytes during growth. Monolayers of FACS-sorted chondrocytes from postnatal day 1-2 (P1-2) mice show faster loss of d2EGFP compared to EGFP, reflecting rapid chondrocyte de-differentiation.

 

Development

A construct in which the mouse Col2a1 promoter/enhancer drives expression of d2EGFP was injected into C57BL/6 x SJL F2 embryos. d2EGFP, derived from plasmid pd2EGFP-N1, carries a modification of EGFP in which residues 422-461 of mouse ornithine decarboxylase (MODC) are fused to the C-terminus of EGFP. This modification creates a destabilized, red-shifted variant of the wildtype GFP which has been optimized for brighter fluorescence and higher expression in mammalian cells. Resultant mice were backcrossed to C57BL/6J for 6 generations by the donating laboratory. 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. One of the 27 markers, on Chromosome 3 was segregating with SJL. Also, 2 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Tg(Col2a1-d2EGFP)1Waho
Name: transgene insertion 1, William A Horton
MGI accession ID: MGI:5904769
Generation method: Transgenic
Attributes: Reporter
Synonyms: Tg(Col2a1-EGFP*)1Waho
Marker symbol: Tg(Col2a1-d2EGFP)1Waho
Marker name: transgene insertion 1, William Horton
Chromosome: UN
Strain of origin: (C57BL/6 x SJL)F2
Expressed genes: GFP,Green Fluorescent Protein,
Site of expression: In the post-natal growth plate, d2EGFP fluorescence is expressed in almost all proliferative chondrocytes and in some hypertrophic chondrocytes.
Molecular note: A construct in which the mouse Col2a1 promoter/enhancer drives expression of d2EGFP was injected into C57BL/6 x SJL F2 embryos. d2EGFP, derived from plasmid pd2EGFP-N1, carries a modification of EGFP in which residues 422-461 of mouse ornithine decarboxylase (MODC) are fused to the C-terminus of EGFP. This modification creates a destabilized, red-shifted variant of the wildtype GFP which has been optimized for brighter fluorescence and higher expression in mammalian cells.