B6.Cg-Gt(ROSA)26Sor^{tm2.1(CAG-Venus,-Sdk1)Jrs}/KrishJ

Stock number: 038966
Product name: RC-sV-Sdk1
Strain synonyms: Rosa-CAG-LOX-STOP-LOX-Venus-3F2A-Sdk1-WPRE
Research areas: Developmental Biology Research, Neurobiology Research, Research Tools, Sensorineural Research

Description

These RC-sV-Sdk1 mice express Venus and Sdk1 after removal of the loxP-flanked STOP cassette via Cre-mediated recombination. These mice are a useful tool for studying interneurons and retinal ganglion cell synapses and their role in visual function.

Detailed description

These RC-sV-Sdk1 mice express (from 5' to 3') a loxP-flanked stop cassette, a Venus reporter sequence, 3F2A, the sidekick cell adhesion molecule 1 (Sdk1) gene sequence, and a WPRE sequence under control of a CAG promoter from the endogenous Gt(ROSA)26Sor locus. SDK1 is a member of the immunoglobulin superfamily that acts as an adhesion molecule that promotes synaptic connectivity in interneurons and retinal ganglion cells (RGCs).

Homozygous mice are viable and fertile. In the absence of Cre, Venus and Sdk1 expression from the Gt(ROSA)26Sor locus is prevented by the STOP cassette. After removal of the loxP-flanked STOP cassette via Cre-mediated recombination, Venus and Sdk1 are initially translated together from the same polycistronic mRNA, and are subsequently separated by the 2A peptide allowing for labeling cells/tissues where cre is expressed. Venus fluorescence is only detectable by immunohistochemisty and no Venus fluorescence is detected prior to Cre recombination.

Of note, RC-sV-Sdk2 mice (Stock No. 038839) are a similar strain with conditional Venus and Sdk2 expression.

Development

A targeting vector was designed with (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP-flanked stop cassette, a Venus reporter sequence, 3F2A (three tandem repeats of foot-and-mouth disease 2A peptide sequence) that mediates ribosomal skipping, the sidekick cell adhesion molecule 1 (Sdk1) gene sequence, a WPRE sequence, and a frt-flanked neomycin resistance (neo) cassette. This targeting vector was inserted into the Gt(ROSA)26Sor locus. The construct was electroporated into (C57BL/6 x 129S4/SvJae)F1-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and resulting chimeric mice were bred with ACTB-Flpe mice on a C57BL/6 congenic background (Stock No. 005703) to remove the neo cassette. Progeny were crossed to remove the FLPe-expressing transgene and resulting RC-sV-Sdk1 mice were bred to C57BL/6J mice for at least 10 generations by the donating laboratory. Upon arrival at The Jackson Laboratory, 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: Gt(ROSA)26Sor^{tm2.1(CAG-Venus,-Sdk1)Jrs}
Name: targeted mutation 2.1, Joshua R Sanes
MGI accession ID: MGI:7546830
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Inserted expressed sequence
Synonyms: RC-sV-Sdk1
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: A targeting vector is designed with (from 5' to 3') a ubiquitously expressed CAG promoter, a loxP-flanked stop cassette, a Venus reporter sequence, 3F2A (three tandem repeats of foot-and-mouth disease 2A peptide sequence) that mediates ribosomal skipping, the sidekick cell adhesion molecule 1 (Sdk1) gene sequence, a WPRE sequence, and an FRT-flanked neomycin resistance (neo) cassette. Flp-mediated recombination removed the neo cassette.