B6.Cg-Gt(ROSA)26Sor^{tm1(CAG-EGFP)Brsy}/J

Stock number: 032290
Product name: MITO-Tag
Research areas: Cancer Research, Neurobiology Research, Research Tools

Description

MITO-Tag mice conditionally express, in a Cre recombinase-dependent manner, a hemagglutinin (HA)-tagged EGFP (codon-optimized) that localizes to mitochondrial outer membranes. These features enable immunocapture of mitochondria from specific cell types without the need for cell sorting techniques.

Detailed description

Mitochondria serve to generate large quantities of cellular energy in the form of adenosine triphosphate (ATP), and perform a number of other functions within eukaryotic cells. Much remains to be learned about their diverse morphologies and mechanisms, however.

Cre recombinase-mediated excision of a loxP-flanked STOP cassette in these MITO-Tag mice enables cell-type-specific expression of hemagglutinin (HA)-tagged EGFP that localizes on the outer membranes of mitochondria. These features enable immunocapture of mitochondria from specific cell types. Mice retaining the floxed-STOP cassette exhibit no overt phenotype.

Development

A construct carrying the following was targeted to intron 1 of the mouse Gt(ROSA)26Sor gene: CAG-loxP-neomycin (reversed)-pA-loxP-3xHA-EGFP-SYNJ2BP-lacZ-BGH-pA. The 3xHA-EGFP-SYNJ2BP components constitute the "MITO-Tag" cassette. SYNJ2BP (synaptojanin 2 binding protein) is also called OMP25, reflecting its Outer Membrane Protein characteristic. The MITO-Tag construct utilizes only a portion of rat Synj2bp/OMP25 (AA109-145), which contains a transmembrane domain and localization signal to the outer mitochondrial membrane. The small fragment of lacZ incorporated in the mutation is not expected to be functional. The genetic modification was created in B6(Cg)-Tyr^c-2J/J-derived CY2.4 embryonic stem (ES) cells and maintained on a C57BL/6J genetic background. The albino (Tyr^c-2J) allele has been bred out of the colony.

Allele

Symbol: Gt(ROSA)26Sor^{tm1(CAG-EGFP)Brsy}
Name: targeted mutation 1, Kivanc Birsoy
MGI accession ID: MGI:6192094
Generation method: Targeted
Attributes: Conditional ready (e.g. floxed); Reporter; Epitope tag
Synonyms: MITO-Tag; Rosa26^{LSL-3XHA-EGFP-OMP25}
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: B6(Cg)-Tyr^c-2J
Molecular note: The targeting construct containing (from 5' t 3') CAG-loxP-neomycin (reversed)-pA-loxP-3xHA-EGFP-SYNJ2BP-lacZ-BGH-pA is targeted to intron 1 of the gene. The 3xHA-EGFP-SYNJ2BP components constitute the "MITO-Tag" cassette targeting the 3XHA epitope tag with GFP to the mitochondrial membrane. The construct includes only the transmembrane domain of the rat Synj2bp/OMP25 (AA109-145) gene. The small fragment of lacZ incorporated in the mutation is not expected to be functional.