B6.129S4-Tg(CAG-EGFP,-Ptgs2,-hrluc)#Hahe/JhewJ

Stock number: 030854
Product name: COX-2 COE
Research areas: Cardiovascular Research, Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Cox-2 COE mice possess a floxed STOP cassette, containing an EGFP sequence, preventing transcription of coding sequences for prostaglandin-endoperoxide synthase 2 (Ptgs2) and humanized Renilla luciferase (hRL) under control of a CAG promoter. In the absence of Cre expression, the transgene constitutively expresses EGFP. Following Cre excision, the transgene expresses Ptgs2 and hRL proteins in a tissue-specific fashion making them useful for studying pain, inflammation, and cancer progression. These mice are useful for repeatedly and noninvasively monitoring Cox-2 overexpression.

Detailed description

Cox-2 COE mice possess a floxed STOP cassette, containing an EGFP sequence, preventing transcription of coding sequences for prostaglandin-endoperoxide synthase 2 (Ptgs2) and humanized Renilla luciferase (hRL). This transgene is under control of a CAG promoter. Ptgs2 encodes Cyclooxygenase-2 (Cox-2), an enzyme that catalyzes the conversion of arachidonic acid to Prostaglandin H (PGH), and eventually to prostanoid lipids. Cox-2 plays a role in thermoregulation, pain perception, inflammation, cardiovascular function, cancer, and neurodegenerative diseases and is a drug target for Cox-2-inhibitors such as non-steroidal anti-inflammatory drugs (NSAIDs). Cox-2 has also been associated with UV-induced tumor development and progression. In the absence of Cre expression, the transgene constitutively expresses EGFP. Following Cre excision, removal of the floxed STOP cassette results in the expression of Ptgs2 and hRL proteins in a tissue-specific fashion. Mice that are homozygous for this allele are viable and fertile.

Development

The Cox-2 COE transgene was designed to contain, from 5’ to 3’, a CAG (CMV enhancer/chicken beta-actin) promoter and a loxP flanked cassette containing an enhanced green fluorescent protein (EGFP) sequence with a polyadenylation signal, a neomycin resistance (neo) cassette, and a translational/transcriptional STOP cassette. Downstream from the second loxP site was is another cassette containing coding sequences for Cox-2 and for humanized Renilla luciferase (hRL), linked by an internal ribosome entry site (IRES). The construct was electroporated into 129S4/SvJae-derived LW1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6 blastocysts and resulting chimeric males were bred with C57BL/6 females. Resulting offspring were further backcrossed to C57BL/6J mice (Stock No. 000664) for 4 more generations. Upon arrival at The Jackson Laboratory Repository, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Tg(CAG-EGFP,-Ptgs2,-hrluc)#Hahe
Name: transgene insertion, Harvey R Herschman
MGI accession ID: MGI:5617514
Generation method: Transgenic
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Marker symbol: Tg(CAG-EGFP,-Ptgs2,-hrluc)#Hahe
Marker name: transgene insertion, Harvey R Herschman
Chromosome: UN
Strain of origin: 129S4/SvJae
Expressed genes: luc,luciferase,firefly; Ptgs2,prostaglandin-endoperoxide synthase 2,mouse, laboratory; GFP,Green Fluorescent Protein,
Site of expression: Elevated expression of Ptgs2 and luciferase was observed in hepatic tissue.
Molecular note: An enhanced green fluorescent protein, a cassette with the SV40 poly-A sequence, a PGK promoter-driven neomycin-bpA cassette, and a translational/transcriptional STOP cassette were inserted in the vector between the CAG promoter and a Ptgs2 cDNA. The coding region for a humanized version of Renilla luciferase (hrluc) was connected with the Ptgs2 coding sequence by an IRES.