Tetracycline regulated transcriptional modulators with altered DNA binding specificities

US5589362A · kind A · utility

Assignees

• BASF SE
• Knoll, Inc. · DE

Inventors

• Hermann Bujard · Heidelberg, DE
• Manfred Gossen · Corona, US
• Wolfgang Hillen · Erlangen, DE
• Vera Helbl · Birkenfeld, DE
• Dirk Schnappinger · Bad Driburg, DE

Key dates

• Filing date: Jun 7, 1995
• Grant date: Dec 31, 1996
• Expiry date: Jun 7, 2015

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2830/32
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Isolated nucleic acid molecules encoding fusion proteins which regulate transcription in eukaryotic cells are disclosed. The fusion proteins of the invention comprise a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells. Methods for regulating transcription of a tet operator-linked gene in a cell are also provided. In one embodiment, the method involves introducing into the cell a nucleic acid molecule encoding a fusion protein which regulates transcription, the fusion protein comprising a Tet repressor having at least one amino acid mutation that confers on the fusion protein an ability to bind a class B tet operator sequence having a nucleotide substitution at position +4 or +6, operatively linked to a polypeptide which regulates transcription in eukaryotic cells, and modulating the concentration of a tetracycline, or analogue thereof, in contact with the cell.