FVB/N-Tg(Pf4-tTA/VP16)42Kra/J

Stock number: 005942
Product name: PF4-tTA-VP16
Research areas: Research Tools

Description

These PF4-tTA-VP16 mice express a transgene containing the tetracycline-controlled transactivator protein (tTA) with its expression driven by the rat platelet factor 4 promoter in megakaryocytes.

Detailed description

Transgenic mice are viable, fertile, normal in size, and do not display any behavioral abnormalities. Transgenic tetracycline-controlled transactivator protein (tTA) expression is limited to megakaryocytes by the rat chemokine (C-X-C motif) ligand 4 (Cxcl4; formerly called platelet factor four (Pf4)) promoter. When bred to mice carrying a gene of interest coupled to a tetracycline-responsive promoter element (TRE; tetO), expression of the target gene is induced specifically in megakaryocytes. Administration of tetracycline/doxycycline sequesters the transgenic protein, and prevents expression of the TRE-regulated target gene.

This mouse originally was designed to be bred with Tg(tetO-Aurkb,lacZ)41Kra (see Stock No. 005941); a transgenic mouse with a bidirectional tetO promoter controlling a mitotic regulator gene (aurora kinase B) and a beta-galactosidase reporter. Megakaryocytes and platelet cells from the bitransgenic offspring show inducible and reversible beta-galactosidase expression. Further, megakaryocytes inducibly express Aurora kinase-B mRNA (but not protein) with a modest effect on megakaryocyte ploidy and no effect on platelet quantity. These mice may be useful in studies of megakaryocyte/platelet lineage cell function, including migration following injury/insult and overexpression of thrombotic agents.

Development

A transgenic construct was designed using the promoter region from rat chemokine (C-X-C motif) ligand 4 (Cxcl4; formerly called platelet factor four (Pf4)) to direct expression of the tetracycline-regulated transactivator, tTA. The construct was injected into fertilized FVB/NJ eggs which were subsequently implanted into pseudopregnant foster mothers. Founder line 42 was obtained and bred to FVB/NJ mice for 3 generations. Transgenic mice were interbred for 5 generations.

Allele

Symbol: Tg(PF4-tTA/VP16)42Kra
Name: transgene insertion 42, Katya Ravid
MGI accession ID: MGI:3656262
Generation method: Transgenic
Attributes: Transactivator
Marker symbol: Tg(PF4-tTA/VP16)42Kra
Marker name: transgene insertion 42, Katya Ravid
Chromosome: UN
Strain of origin: FVB/NJ
Expressed genes: tTA,tetracycline-controlled transactivator,E. coli
Site of expression: Expresses tTA in megakaryocytes.
Molecular note: The rat platelet factor 4 (RP4) promoter was cloned into the tTA-containing plasmid, replacing the existing CMV promoter. The promoter will drive expression of the transactivator tTA fused to the viral coactivator VP16. Expression is repressed with administration of doxycycline and is induced by removal of doxycycline from the diet of the mice.The transgene also contains 1.7 kb of the 3'end of the human beta-globin gene including the 3' intron and polyadenylation signal to permit stable RNA production in mice. A stop codon in the transgene prevents translation of beta-globin into the protein.