Antibiotic-based gene regulation system

A mammalian cell comprising a nucleic acid, said nucleic acid comprising an ETR sequence operatively linked to a first eukaryotic promoter.

A method for regulating expression of an ETR-linked gene in a mammalian cell, comprising introducing into the cell a nucleic acid molecule encoding an ETR-binding polypeptide, thereby rendering the ETR-linked gene capable of regulation by an antibiotic that binds to the ETR-binding polypeptide.

A process for producing a protein comprising:

• a) culturing a mammalian cell containing an ETR-linked gene that encodes the protein and a nucleic acid molecule encoding an ETR-binding polypeptide, and
• b) modulating the level of an antibiotic that contacts the cell thereby regulating expression of the ETR-linked gene.

A method for screening for a candidate antibiotic or immunomodulatory compound, the method comprising contacting a mammalian cell with a test compound, wherein the mammalian cell comprises a polypeptide capable of binding an ETR sequence in the absence of its cognate antibiotic and a nucleotide sequence encoding a reporter gene operatively linked to an ETR sequence, and wherein a change in the transcription of the reporter gene indicates that the test compound is a candidate antibiotic or immunomodulatory compound.

A method for screening for a candidate antibiotic or immunomodulatory compound comrpising the steps of:

• a) contacting a test compound with a polynucleotide comprising an ETR and a polypeptide capable of binding an ETR sequence, wherein the polynucleotide is immobilized on a solid support;
• and
• b) assaying for release of the polypeptide from the solid support, wherein release of the polypeptide indicates that the test compound is a candidate antibiotic or immunomodulatory compound.

A method for screening for a candidate antibiotic or immunomodulatory compound comprising the steps of:

• a) contacting a test compound with a polynucleotide comprising an ETR and a polypeptide capable of binding an ETR sequence, wherein the polypeptide is immobilized on a solid support;
• and
• b) assaying for release of the polynucleotide from the solid support, wherein release of the polynucleotide indicates that the test compound is a candidate antibiotic or immunomodulatory compound.

A viral vector comprising a polynucleotide encoding a polypeptide capable of binding an ETR sequence in the absence of its cognate antibiotic.

A transgenic mouse having cells comprising a polynucleotide encoding a polypeptide capable of binding an ETR sequence in the absence of its cognate antibiotic.

• antibiotic.

A transgenic mouse having cells comprising a polynucleotide, said polynucleotide comprising an ETR sequence operatively linked to a first eukaryotic promoter.

A kit comprising a first polynucleotide encoding a polypeptide capable of binding an ETR sequence in the absence of its cognate antibiotic and a second polynucleotide comprising a promoter operably linked to an ETR sequence.

A kit comprising:

• a) a eukaryotic cell comprising a first polynucleotide that encodes a polypeptide capable of binding an ETR sequence in the absence of its cognate antibiotic;
• and
• b) a second polynucleotide comprising a promoter operably linked to an ETR sequence.

A plant cell comprising a nucleic acid encoding a polypeptide capable of activating or repressing an ETR sequence in the absence of its cognate antibiotic.

A plant cell comprising a nucleic acid, said nucleic acid comprising an ETR sequence operatively linked to a first eukaryotic promoter.

A method for regulating expression of an ETR-linked gene in a plant cell, comprising introducing into the cell a nucleic acid molecule encoding an ETR-binding polypeptide, thereby rendering the ETR-linked gene capable of regulation by an antibiotic that binds to the ETR-binding polypeptide.

A fusion polypeptide comprising a polypeptide capable of binding an ETR sequence operatively linked to a second polypeptide capable of activating or repressing transcription in eukaryotic cells.