B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J

Stock number: 008344
Product name: TetTag
Research areas: Neurobiology Research, Research Tools

Description

The TetTag mouse is a bi-transgenic mutant that has tetracycline (or tetracycline analog) inducible expression of beta-galactosidase in activated neurons. This strain provides a tool that allows the inducible stable labeling of activated neurons.

Detailed description

The TetTag mouse is a bi-transgenic mutant that has tetracycline (or a tetracycline analog such as doxycycline) inducible expression of beta-galactosidase in activated neurons. Two independently generated transgenic strains were crossed to produce this bi-transgenic TetTag strain. In the first transgenic construct, the tetracycline-controlled transactivator (tTA) protein and a two hour half-life Green Fluorescent Protein (shEGFP) are expressed under the direction of the fos, FBJ osteosarcoma oncogene, minimal promoter. The second transgenic construct expresses a nuclear-localizing beta-galactosidase gene and the doxycycline insensitive tetracycline regulated transactivator (containing point mutation, H100Y), under the control of the tetO, tetracycline-responsive regulatory element.

Doxycycline administration prevents expression beta-galactosidase in neurons. In the absence of doxycycline, tau-LacZ is induced in neurons of the dentate gyrus (DG) and CA1 neuron, with most of the LacZ-positive neurons displaying typical morphologies of granule cells and pyramidal neurons in the DG and CA1, respectively. Neurons that were induced to express beta-galactosidase during the absence of doxycycline administration will continue to express beta-galactosidase via the doxycycline insensitive mutant tTA^H100Y. Mice hemizygous for each transgene are viable and fertile. The strain is maintained with doxycycline administration to prevent neuronal beta-galactosidase expression. The donating investigator has not attempted to make these mice homozygous for the transgenes. This strain provides a tool that allows the inducible stable labeling of activated neurons.

Of note, cfos-htTA mice (cfos-tTA/cfos-shEGFP) harboring only the co-injected cfos-tTA/cfos-shEGFP transgenes are available at The Jackson Laboratory as Stock No. 018306.

Development

To produce this bi-transgenic line, two independently generated single transgenic lines were crossed.

The first transgenic line was generated by co-injection of a construct containing the tetracycline regulated transactivator (tTA), SV40 polyadenylation signal sequence and the fos, FBJ osteosarcoma oncogene, minimal promoter and a second construct containing a two hour half-life Green Fluorescent Protein under the control of the fos, FBJ osteosarcoma oncogene, minimal promoter, into fertilized B6D2F2 mouse eggs. These co-injected transgenes segregated as a single transgene.

The second transgenic line was generated using a construct including sequence encoding tau-LacZ and the doxycycline insensitive tetracycline regulated transactivator (containing point mutation, H100Y), under the control of the tetO, tetracycline-responsive regulatory element. This construct was injected into fertilized B6D2F2 mouse eggs.

The two independent transgenic lines were then crossed to produce this bi-transgenic strain. The mice were then backcrossed to C57BL/6J for 3 generations.

Allele

Symbol: Tg(tetO-lacZ,tTA*)1Mmay
Name: transgene insertion 1, Mark Mayford
MGI accession ID: MGI:3818450
Generation method: Transgenic
Attributes: Reporter; Inducible; Transactivator
Marker symbol: Tg(tetO-lacZ,tTA*)1Mmay
Marker name: transgene insertion 1, Mark Mayford
Chromosome: UN
Strain of origin: (C57BL/6 x DBA/2)F2
Expressed genes: tTA,tetracycline-controlled transactivator,E. coli; lacZ,beta-galactosidase,E. coli
Site of expression: activated neurons in the basolateral amygdala
Molecular note: A construct including the sequence encoding tau-LacZ and a doxycycline-insensitive tetracycline regulated transactivator (containing point mutation, H100Y), under the control of a bi-cistronic tetO promoter (tetracycline-responsive regulatory element) was injected into fertilized eggs.

Allele

Symbol: Tg(Fos-tTA,Fos-EGFP*)1Mmay
Name: transgene insertion 1, Mark Mayford
MGI accession ID: MGI:3818447
Generation method: Transgenic
Attributes: Reporter; Transactivator
Marker symbol: Tg(Fos-tTA,Fos-EGFP*)1Mmay
Marker name: transgene insertion 1, Mark Mayford
Chromosome: UN
Strain of origin: (C57BL/6 x DBA/2)F2
Expressed genes: tTA,tetracycline-controlled transactivator,E. coli; GFP,Green Fluorescent Protein,
Site of expression: activated neurons in the basolateral amygdala
Molecular note: A construct containing the Fos (FBJ osteosarcoma oncogene, minimal promoter) driving the tetracycline regulated transactivator (tTA) sequence, the SV40 polyadenylation signal sequence and a second construct containing a two hour half-life Green Fluorescent Protein (shEGFP) under the control of the Fos minimal promoter, were co-injected into fertilized B6D2F2 mouse eggs. These co-injected transgenes segregate as a single transgene.