Mutant Receptors and Their Use in a Nuclear Receptor-Based Inducible Gene Expression System
AssigneeIntrexon Corporation
InventorsSubba Reddy PALLI, Mohan Basavaraju KUMAR
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Abstract
This invention relates to the field of biotechnology or genetic engineering. Specifically, this invention relates to the field of gene expression. More specifically, this invention relates to novel substitution mutant receptors and their use in a nuclear receptor-based inducible gene expression system and methods of modulating the expression of a gene in a host cell for applications such as gene therapy, large scale production of proteins and antibodies, cell-based high throughput screening assays, functional genomics and regulation of traits in transgenic organisms.
Claims35
-21. (canceled)
A recombinant gene expression modulation system comprising a recombinant gene expression cassette that is capable of being expressed in a host cell, the gene expression cassette comprising a polynucleotide that encodes a first polypeptide comprising:
- (a) a transactivation domain;
- (b) a DNA-binding domain that recognizes a response element associated with a gene whose expression is to be modulated;
- and
- (c) a nuclear receptor ligand binding domain that is capable of binding a diacylhydrazine,
- wherein said nuclear receptor ligand binding domain comprises either an amino acid residue substitution mutation at amino residue 128 of SEQ ID NO:
- 1, or an amino acid residue substitution mutation at each of amino acid residues 110 and 128 of SEQ ID NO:
- 1.
A recombinant gene expression modulation system comprising:
- (a) a first recombinant gene expression cassette that is capable of being expressed in a host cell comprising a polynucleotide that encodes a first polypeptide comprising:
- (i) a DNA-binding domain that recognizes a response element associated with a gene whose expression is to be modulated;
- and
- (ii) a first nuclear receptor ligand binding domain;
- and
- (b) a second recombinant gene expression cassette that is capable of being expressed in the host cell comprising a polynucleotide that encodes a second polypeptide comprising:
- (i) a transactivation domain;
- and
- (ii) a second nuclear receptor ligand binding domain,
- wherein one of said first and said second nuclear receptor ligand binding domains is capable of binding a diacylhydrazine, and comprises either an amino acid residue substitution mutation at amino residue 128 of SEQ ID NO:
- 1, or an amino acid residue substitution mutation at each of amino acid residues 110 and 128 of SEQ ID NO:
- 1.
An isolated polynucleotide encoding a nuclear receptor ligand binding domain that is capable of binding a diacylhydrazine,
- wherein said nuclear receptor ligand binding domain comprises either an amino acid residue substitution mutation at amino residue 128 of SEQ ID NO:
- 1, or an amino acid residue substitution mutation at each of amino acid residues 110 and 128 of SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.