Polynucleotide
AssigneeMillipore Corporation
InventorsMichael Antoniou, Robert Crombie
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Abstract
The present invention relates to a polynucleotide comprising a ubiquitous chromatin opening element (UCOE). The present invention also relates to a vector comprising the polynucleotide sequence, a host cell comprising the vector, use of the polynucleotide, vector or host cell in therapy and in an assay, and a method of identifying UCOEs. The UCOE opens chromatin or maintains chromatin in an open state and facilitates reproducible expression of an operably-linked gene in cells of at least two different tissue types.
Claims52
An isolated polynucleotide comprising
- a) an element comprising an extended methylation-free CpG-island;
- b) an expressible gene, wherein said expressible gene is operably-linked to said CpG-island;
- and
- c) a tissue-specific promoter, operably-linked to said gene, wherein said promoter is not naturally operably-linked to said CpG-island,
- wherein said element facilitates activation of transcription of said gene.
An isolated polynucleotide comprising
- a) an element comprising an extended methylation-free CpG-island comprising at least one endogenous promoter;
- b) an expressible gene, wherein said expressible gene is operably-linked to said CpG-island, further wherein said expressible gene is not naturally linked to said CpG-island;
- and
- c) a further promoter, external to said CpG-island, operably-linked to said gene, wherein said further promoter is a tissue-specific promoter not naturally operably-linked to said CpG-island,
- wherein said element facilitates activation of transcription of said gene.
A vector comprising an isolated polynucleotide comprising
- a) an element comprising an extended methylation-free CpG-island, wherein any DNAse I hypersensitive sites in said element are associated with an endogenous promoter;
- b) a multiple cloning site operably-linked to said CpG-island, into which an expressible gene can be cloned;
- and
- c) a further promoter operably-linked to said multiple cloning site, wherein said further promoter is a tissue-specific promoter not naturally operably-linked to said CpG-island.
An isolated polynucleotide comprising
- a) an element comprising an extended methylation-free CpG-island comprising at least one endogenous promoter;
- b) an expressible gene, wherein said expressible gene is operably-linked to said CpG-island, further wherein said expressible gene is not naturally linked to said CpG-island;
- and
- c) a further promoter, external to said CpG-island, operably-linked to said gene, wherein said further promoter is a tissue-specific promoter not naturally operably-linked to said CpG-island,
- wherein said element facilitates activation of transcription of said gene.
Dependencies are inferred from the claim text.