Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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Stem Cell Related Patent Number US5789156

Title:Tetracycline-regulated transcriptional inhibitors
Inventors:Bujard, Hermann; Heidelberg, Germany
Gossen, Manfred; El Cerrito, CA, USA
Summary:This invention describes novel tetracycline-regulated transcription modulators comprising fusion proteins which bind to tet operator sequences to activate or inhibit transcription. Described herein is an isolated nucleic acid encoding a fusion protein which inhibits transcription in eukaryotic cells, the fusion protein comprising a first polypeptide which binds to tet operator sequences, operatively linked to a heterologous second polypeptide which inhibits transcription in eukaryotic cells. Further described are nucleic acid molecules and proteins considered to be useful for regulating the expression of genes in eukaryotic cells and organisms in a highly controlled manner. The internal regulatory system of this invention provides for the transcription of a tet operator-linked nucleotide sequence which is inhibited by a transcriptional inhibitor fusion protein composed of two polypeptides: the first polypeptide binds to tet operator sequences either in the absence but not the presence of tetracycline or an analogue thereof, or in the presence but not the absence of tetracycline or an analogue thereof, and the second polypeptide directly or indirectly inhibits transcription in eukaryotic cells. Further disclosed are methods by which the fusion protein comprises a tet repressor operatively linked to a transcriptional silencer polypeptide, as well as a fusion protein comprising a mutated tet repressor operatively linked to a transcriptional silencer polypeptide. The fusion proteins disclosed herein are useful for reducing the level of transcription of a tet operator-linked target gene, and they can also be used in combination with tetracycline-regulated transcriptional activator fusion proteins to allow for precise regulation of the expression of one or multiple target genes. Further provided herein are kits including the components of the regulatory system of the invention.
Abstract:Nucleic acid molecules and proteins useful for regulating the expression of genes in eukaryotic cells and organisms in a highly controlled manner are disclosed. In the regulatory system of the invention, transcription of a tet operator-linked nucleotide sequence is inhibited by a transcriptional inhibitor fusion protein composed of two polypeptides, a first polypeptide which binds to tet operator sequences either (i) in the absence but not the presence of tetracycline (or an analogue thereof) or (ii) in the presence but not the absence of tetracycline (or an analogue thereof), and a second polypeptide which directly or indirectly inhibits transcription in eukaryotic cells. In one embodiment, the fusion protein comprises a Tet repressor operatively linked to a transcriptional silencer polypeptide. In another embodiment, the fusion protein comprises a mutated Tet repressor operatively linked to a transcriptional silencer polypeptide. The fusion proteins of the invention are useful for reducing the level of transcription of a tet operator-linked target gene. Moreover, the fusion proteins of the invention can be used in combination with tetracycline-regulated transcriptional activator fusion proteins to allow for precise regulation of the expression of one or multiple target genes. Kits including the components of the regulatory system of the invention are also encompassed by the invention.
US Patent Website:Click Here for Full Text of Patent
Title Number:US5789156
Application Number:US1995000383754
Date Filed:03/02/1995
Date Published:04/08/1998
Assignee:BASF AG, Ludwigshafen, Germany Knoll AG, Ludwigshafen, Germany


 
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