PhAMexcised (photo-activatable mitochondria) mice express a mitochondrial-specific version of Dendra2 green/red photoswitchable monomeric fluorescent protein in mitochondria. Exposure of mitochondria to 405 nm laser light irreversibly switches green fluorescence to red, enabling measurements of mitochondrial fusion and transport.
David C Chan, California Institute of Technology
Genetic Background | Generation |
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|
Allele Type | Gene Symbol | Gene Name |
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Targeted (Reporter) | Gt(ROSA)26Sor | gene trap ROSA 26, Philippe Soriano |
Dendra2 is a green-to-red, irreversibly photoswitchable monomeric fluorescent protein derived from octocoral Dendronephthya sp. Its fusion with the mitochondrial targeting signal of subunit VIII of cytochrome c oxidase generates "mito-Dendra2", capable of mitochondrial-specific fluorescence.
In this PhAMexcised strain, mito-Dendra2 is introduced into the ubiquitously-expressed Gt(ROSA)26Sor gene downstream of a CAG (CMV-βactin) promoter. All organs isolated from these animals (e.g. central nervous system, skeletal muscle, heart, testis, lung, kidney, thymus) exhibit bright green fluorescence localized specifically to the mitochondrial compartment. Homozygotes are viable and fertile.
When a sub-population of mitochondria is exposed to 405 nm laser light, the green signal photo-converts to red fluorescence, enabling measurements of mitochondrial fusion (indicated by mixing of green and red signals) and transport.
Dendra2 (a green to red irreversibly photoswitchable monomeric fluorescent protein derived from octocoral Dendronephthya sp.) fused to the mitochondrial targeting signal of subunit VIII of cytochrome c oxidase (mito-Dendra2), bovine growth hormone (bGH) polyadenylation signal, and an FRT-flanked neomycin cassette were introduced to Gt(ROSA)26Sor downstream of a CAG (CMV-βactin) promoter and loxP-flanked termination signal. The mutation was created in TC1 129S6/SvEvTac-derived embryonic stem cells. Crosses with a C57BL/6 background FLP strain excised the FRT-flanked neomycin cassette. Crosses with a germline cre deleter strain (Meox2-cre) excised the remaining floxed termination cassette. This strain was maintained on a mixed C57BL/6-129 genetic background by the donating lab.
Expressed Gene | Dendra, Dendra photoswitchable fluorescent protein, octocoral Dendronephthya |
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Site of Expression | Cre excision of the stop signal results in expression of Dendra2, a green-to-red irreversibly photo-switchable fluorescent protein, in the mitochondria of cre-expressing tissues. |
Allele Name | targeted mutation 1.1, David C Chan |
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Allele Type | Targeted (Reporter) |
Allele Synonym(s) | Gt(ROSA)26Sortm1.1(CAG-Mito-Dendra2)Dcc; mito-Dendra2; PhAMexcised |
Gene Symbol and Name | Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano |
Gene Synonym(s) | |
Promoter | CAG, CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, |
Expressed Gene | Dendra, Dendra photoswitchable fluorescent protein, octocoral Dendronephthya |
Site of Expression | Cre excision of the stop signal results in expression of Dendra2, a green-to-red irreversibly photo-switchable fluorescent protein, in the mitochondria of cre-expressing tissues. |
Strain of Origin | 129S6/SvEvTac |
Chromosome | 6 |
Molecular Note | Dendra2 (a green to red irreversibly photoswitchable monomeric fluorescent protein derived from octocoral Dendronephthya sp.) fused to the mitochondrial targeting signal of subunit VIII of cytochrome c oxidase (mito-Dendra2), bovine growth hormone (bGH) polyadenylation signal, and an FRT-flanked neomycin cassette were introduced to Gt(ROSA)26Sor downstream of a CAG (CMV-beta-actin) promoter and loxP-flanked termination signal. Flp-mediated recombination removed the neo cassette and cre-mediated recombination excised the floxed termination cassette. |
Mutations Made By | David Chan, California Institute of Technology |
Homozygotes and heterozygotes are viable and fertile.
When using the PhaM excised mouse strain in a publication, please cite the originating article(s) and include JAX stock #018397 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Heterozygous for Gt(ROSA)26Sor<tm1.1(CAG-COX8A/Dendra2)Dcc> |
Frozen Mouse Embryo | B6;129S-Gt(ROSA)26Sor<tm1.1(CAG-COX8A/Dendra2)Dcc>/J | $2595.00 |
Frozen Mouse Embryo | B6;129S-Gt(ROSA)26Sor<tm1.1(CAG-COX8A/Dendra2)Dcc>/J | $2595.00 |
Frozen Mouse Embryo | B6;129S-Gt(ROSA)26Sor<tm1.1(CAG-COX8A/Dendra2)Dcc>/J | $3373.50 |
Frozen Mouse Embryo | B6;129S-Gt(ROSA)26Sor<tm1.1(CAG-COX8A/Dendra2)Dcc>/J | $3373.50 |
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