STOCK Gt(ROSA)26Sor^{tm3.2(CAG-HRP)Ddg}/J

Stock number: 034750
Product name: ROSA26^FSF-ER-HRP
Research areas: Research Tools

Description

ROSA26^FSF-ER-HRP mice conditionally express endoplasmic reticulum (ER)-targeted horse radish peroxidase (HRP) in a cre recombinase-dependent manner. Flp recombinase-mediated excision of an FRT-flanked Stop cassette enables directed expression of horse radish peroxidase fused to an endoplasmic reticulum retention signal, useful in multiplexed electron microscopy labeling.

The donating laboratory describes this strain as an ER-ERP strain: horseradish peroxidase fused to immunoglobulin kappa light chain leader sequence and KDEL endoplasmic reticulum retention signal.

Detailed description

ROSA26^FSF-ER-HRPmice conditionally express endoplasmic reticulum (ER)-targeted horse radish peroxidase (HRP) in a Flp recombinase-dependent manner. Flp recombinase-mediated excision of an FRT-flanked Stop cassette enables directed expression of horse radish peroxidase fused to an endoplasmic reticulum retention signal, useful in multiplexed electron microscopy labeling.

Expression has been observed by the donating laboratory in all tissues (cortex, spinal cord and dorsal root ganglion (DRG)) where the Stop cassette has been excised using various cre recombinases.

Strains carrying either dual recombinase loxP- and FRT-flanked Stop cassettes (see Stock No. 034748, ROSA26^DR-ER-HRP) or the loxP-flanked Stop cassette (see Stock No. 034746, ROSA26^LSL-ER-HRP) regulating ER-HRP expression are also available.

Similarly-constructed conditional mutant strains expressing HRP fused to synaptophysin (SV-HRP), labeling the lumen of synaptic vesicles, have also been created:
Stock No. 034749, ROSA26^DR-SV-HRP: SV-HRP expression regulated by the concurrent excision of both loxP- and FRT-flanked Stop cassettes.
Stock No. 034747, ROSA26^LSL-SV-HRP: SV-HRP expression regulated by the excision of a single loxP-flanked Stop cassette.
Stock No. 034745, ROSA26^FSF-SV-HRP: SV-HRP expression regulated by the excision of a single FRT-flanked Stop cassette.

Development

The donating laboratory describes this strain as an ER-ERP strain: horseradish peroxidase fused to immunoglobulin kappa light chain leader sequence and KDEL endoplasmic reticulum retention signal.

ROSA26^DR-ER-HRP mice were created by replacing the tdTomato cassette in the Ai65 targeting vector (see Stock No. h) with DR-ER-HRP, a dual (Cre and Flp) recombinase (DR)-dependent, endoplasmic reticulum (ER)-targeted, horse radish peroxidase (HRP) reporter cassette.

The following were introduced to intron 1 of the mouse Gt(ROSA)26Sor gene via homologous recombination in 129S4/SvJae-derived J1 embryonic stem (ES) cells: CAG-FRT-Stop-FRT-loxP-Stop-loxP-ER ("KDEL" target peptide for endoplasmic reticulum retention)-HRP (horse radish peroxidase)-WPRE (Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element)-bGHpA (bovine growth hormone polyadenylation signal)-attB-PGK-NeoR-PGKpA-attP.

ROSA26^DR-ER-HRP mice were crossed with a germline cre recombinase strain to excise the loxP-Stop-loxP component of the construct, leaving the FRT-Stop-FRT feature found in this ROSA26^FSF-ER-HRP allele.

This strain was maintained on a mixed genetic background including C57BL/6, 129S4/SvJae and CD1 by the donating laboratory.

Allele

Symbol: Gt(ROSA)26Sor^{tm3.2(CAG-HRP)Ddg}
Name: targeted mutation 3.2, David D Ginty
MGI accession ID: MGI:6385524
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter
Synonyms: ROSA26^FSF-ER-HRP
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: 129S4/SvJae
Molecular note: The targeting vector is created by replacing the tdTomato cassette in Gt(ROSA)26Sor^{tm65.1(CAG-tdTomato)Hze} with DR-ER-HRP, a dual (Cre and Flp) recombinase (DR)-dependent, endoplasmic reticulum (ER)-targeted, horse radish peroxidase (HRP) reporter cassette. The following was introduced to intron 1 of the mouse Gt(ROSA)26Sor gene - CAG-FRT-Stop-FRT-loxP-Stop-loxP-ER ("KDEL" target peptide for endoplasmic reticulum retention)-HRP (horse radish peroxidase)-WPRE (Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element)-bGHpA (bovine growth hormone polyadenylation signal)-attB-PGK-NeoR-PGKpA-attP. Cre-mediated recombination removed the floxed stop cassette.