US 5,859,310 A · Mice transgenic for a tetracycline-controlled transcriptional activator

Abstract

Transgenic mice carrying two transgenes, the first coding for a transactivator fusion protein comprising a tet repressor and a polypeptide which directly or indirectly activates transcription of a tet operator-linked gene in eucaryotic cells, and the second comprising a gene operably linked to a minimal promotor operably linked to at least one tet operator sequence, are disclosed. Isolated DNA molecules (e.g., targeting vectors) for integrating a polynucleotide sequence encoding a transactivator of the invention at a predetermined location within a second target DNA molecule by homologous recombination are also disclosed. Transgenic mice having the DNA molecules of the invention integrated at a predetermined location in a chromosome by homologous recombination are also encompassed by the invention. Methods to regulate the expression of a tet operator linked-gene of interest by administering tetracycline or a tetracycline analogue to a mouse of the invention are also disclosed. The regulatory system of the invention allows for conditional inactivation or modulation of expression of a gene of interest in a host cell or mouse.

Claims20

Dependent claims

A transgenic mouse having a transgene integrated into the genome of the mouse and also having a tet operator-linked gene in the genome of the mouse, wherein:

the transgene comprises a transcriptional regulatory element functional in cells of the mouse operatively linked to a polynucleotide sequence encoding a tetracycline-controllable transactivator fusion protein (tTA),
said fusion protein comprises a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription of said tet operator-linked gene in eucaryotic cells,
said tet operator-linked gene confers a detectable and functional phenotype on the mouse when expressed in cells of the mouse,
said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fission protein that are sufficient to activate transcription of the tet operator-linked gene;
and
in the absence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tet operator-linked gene and activates transcription of the tet operator linked gene such that the tet operator-linked gene is expressed at a level sufficient to confer the detectable and functional phenotype on the mouse, wherein the level of expression of tet operator-linked gene can be downmodulated by administering tetracycline or a tetracycline analogue to the mouse.

3 more elements

A transgenic mouse having a transgene integrated into the genome of the mouse, wherein:

the transgene comprises a polynucleotide sequence encoding a tetracycline-controllable transactivator fusion protein (tTA) and a tTA-responsive promoter,
said fusion protein comprises a Tet repressor operably linked to a polypeptide which directly or indirectly activates transcription of a gene of interest in eucaryotic cells,
the transgene is integrated by homologous recombination at a predetermined location within a said gene of interest within cells of the mouse such that expression of the fission protein is controlled by 5' regulatory elements of the gene of interest and expression of the gene of interest is controlled by the tTA-responsive promoter,
expression of the gene of interest confers a detectable and functional phenotype on the mouse,
said transgene is expressed in cells of the mouse at a level sufficient to produce amounts of said fusion protein that are sufficient to activate transcription of the gene of interest linked to the tTA-responsive promoter, and
in the absence of tetracycline or a tetracycline analogue in the mouse, said fusion protein binds to the tTA-responsive promoter and activates transcription of the gene of interest such that the gene of interest is expressed at a level sufficient to confer the detectable and functional phenotype on the mouse, wherein the level of expression of the gene of interest can be downmodulated by administering tetracycline or a tetracycline analogue to the mouse.

3 more elements

Dependencies are inferred from the claim text.

Mice transgenic for a tetracycline-controlled transcriptional activator

Life of this patent

Mice transgenic for a tetracycline-controlled transcriptional activator

Life of this patent

Abstract

Claims20