STOCK Tg(CAG-mKate2)1Poche/J

Stock number: 032188
Product name: mito::mKate2
Research areas: Neurobiology Research, Research Tools

Description

These mito::mKate2 mice are useful for widespread in vivo and ex vivo imaging of mitochondrial dynamics within living tissues.

Detailed description

The mito::mKate2 transgenic mice have the ubiquitous CAG promoter directing expression of a far-red fluorescent protein (mKate2) with N-terminal fusion of the cytochrome c oxidase subunit VIII mitochondrial targeting signal, resulting in direct fluorescence in the mitochondria. mKate2 mice can be used for in vivo and ex vivo imaging of mitochondria within living tissues. Fluorescence is detected in the tissues tested, specifically, adult flexor digitorum brevis (FDB) muscle, sperm midpiece, Purkinje neurons of the cerebellum, retina, heart, and liver. mito::mKate2 are useful for monitoring mitochondrial dynamics coincident with cell cycle kinetics during mouse embryonic development and in adult tissues. mKate2 can be visualized with other traditional fluorescent reporters (such as GFP, YFP, CFP, and DsRed) since its fluorescence lies outside of the blue/green/red color range. Hemizygous mice are viable and fertile, the donating lab has not attempted to make a homozygous colony.

Development

The mito::mKate2 transgene is composed of an N-terminal mitochondrial targeting pre-sequence (derived from human cytochrome c oxidase subunit VIII [COX8A]), fused to a far-red fluorescent protein, mKate2. The mito::mKate2 fusion protein cDNA sequence was placed downstream of the CMV enhancer/chicken beta-actin (CAG) promoter. The transgene was microinjected into C57BL/6J (Stock No. 000664) fertilized eggs. Transgenic mice were bred with CD1 outbred mice, and founder line 1 was characterized. The mito::mKate2 transgenic colony was maintained by backcrossing to C57BL/6J mice for at least 2 generations prior to sending males to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To generate our live colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6J mice.

Allele

Symbol: Tg(CAG-mKate2)1Poche
Name: transgene insertion 1, Ross Poche
MGI accession ID: MGI:6161749
Generation method: Transgenic
Attributes: Reporter
Synonyms: mito::mKate2
Marker symbol: Tg(CAG-mKate2)1Poche
Marker name: transgene insertion 1, Ross Poche
Chromosome: UN
Strain of origin: C57BL/6J
Expressed genes: RFP,Red Fluorescent Protein,coral
Site of expression: Mitochondria
Molecular note: The transgene is composed of an N-terminal mitochondrial targeting pre-sequence (derived from human cytochrome c oxidase subunit VIII [COX8A]), fused to a far-red fluorescent protein, mKate2. The fusion protein cDNA sequence was placed downstream of the CMV enhancer/chicken beta-actin (CAG) promoter.