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US 2003/0165903 A1Published application
Chimeric histone acetyltransferase polypeptides
AssigneeCeres, Inc.
InventorsVan-Dinh Dang, Jack Okamuro
Life of this patent
Estimating the term…
Abstract
Chimeric polypeptides are disclosed that comprise a first polypeptide segment having histone acetyltransferase enzymatic activity and a second polypeptide segment that is similar to a subunit of a chromatin-associated histone deacetyltransferase protein complex. Also disclosed are nucleic acids encoding such chimeric polypeptides and eukaryotic organisms expressing such chimeric polypeptides.
Claims100
A chimeric polypeptide comprising:
- a. a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- b. a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is linked to a terminus of said first polypeptide segment via at least one covalent bond.
1 more element
A eukaryotic organism comprising a nucleic acid encoding a chimeric polypeptide, said chimeric polypeptide comprising:
- a. a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- b. a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment.
1 more element
A eukaryotic organism comprising:
- a. a first nucleic acid construct comprising a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
- i) a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment;
- and
- b. a second nucleic acid construct comprising a second promoter conferring cell type-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element.
4 more elements
A method for detecting the expression of one or more genes in a eukaryote, said method comprising:
- a. isolating macromolecules from one or more specific cells in said eukaryote, said eukaryote comprising a nucleic acid construct that has a promoter operably linked to a coding sequence that encodes:
- i) a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment;
- and
- b. determining the presence or amount of at least one of said macromolecules in at least one of said specific cells.
4 more elements
A method for detecting the expression of one or more genes in a eukaryote, said method comprising:
- a. isolating macromolecules from one or more specific cells in said eukaryote, said eukaryote comprising:
- i) a first nucleic acid construct having a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
- a) a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- b) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment;
- and
- ii) a second nucleic acid construct comprising a second promoter conferring cell type-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element;
- and
- b. determining the presence or amount of at least one of said macromolecules in at least one of said specific cells.
7 more elements
A method for modulating gene expression in a eukaryote, said method comprising making a eukaryote having a nucleic acid construct comprising a cell-type specific promoter operably linked to a coding sequence that encodes:
- a. a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- b. a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment;
- and
- wherein said eukaryote exhibits modulated gene expression in cells in which said promoter confers cell-type specific transcription.
3 more elements
A method for modulating gene expression in a eukaryote, said method comprising making a eukaryote, said eukaryote comprising:
- a. a first nucleic acid construct having a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
- i) a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment;
- and
- b) a second nucleic acid construct comprising a second promoter conferring cell type-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element,
- wherein said eukaryote exhibits modulated gene expression in cells in which said second promoter confers cell-type specific expression.
5 more elements
A method for making a genetically modified eukaryote, said method comprising:
- a. providing a first eukaryote comprising a first nucleic acid construct, said first nucleic acid construct comprising a first promoter and a transcription activator element operably linked to a coding sequence, said coding sequence encoding:
- i) a first polypeptide segment that exhibits histone acetyltransferase activity;
- and
- ii) a second polypeptide segment, wherein said second polypeptide segment has 40% or greater sequence identity to a subunit of a histone deacetylase chromatin-associated protein complex,
- wherein a terminus of said second polypeptide segment is covalently linked to a terminus of said first polypeptide segment;
- and
- b. providing a second eukaryote comprising a second nucleic acid construct, said second nucleic acid construct comprising a second promoter conferring embryo-specific transcription, said second promoter operably linked to a coding sequence for a polypeptide that binds said transcription activator element,
- c. crossing said first eukaryote and said second eukaryote to form genetically modified progeny that are sterile.
5 more elements
Dependencies are inferred from the claim text.