Method of detecting Polycomb Repressive Complex activity
AssigneeImba—Institut FüR Molekulare Biotechnologie GmbH
InventorsOliver Bell, Hagar Moussa, Ramesh Yelagandula
Life of this patent
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Abstract
A method of detecting Polycomb Repressive Complex (PRC) activity in a cell providing a cell with a DNA having a protein binding site and at least one reporter gene expression site is operatively connected to the protein binding site, and with a DNA containing a recombinant gene of a binding protein, the binding protein being capable of binding to the protein binding site, wherein the binding protein is fused to a member of the PRC, the method including the step of expressing the recombinant gene, letting the fused binding protein bind to the protein binding site and detecting at least one reporter gene expression.
Claims25
A method of detecting Polycomb Repressive Complex (PRC) activity in a vertebrate cell, the method comprising the step of providing vertebrate cells with a DNA having a protein binding site and at least one reporter gene expression site that is operatively connected to said protein binding site, wherein said protein binding site is located at a transcriptionally neutral locus or region of the genome of the vertebrate cells, wherein the transcriptionally neutral locus or region lacks active and repressive histone modifications, and the vertebrate cells comprise a DNA comprising a recombinant gene of a binding protein, said binding protein being capable of binding to said protein binding site, wherein said binding protein is fused to a member of the PRC, said method further comprising the steps of expressing said recombinant gene, letting said fused binding protein bind to said protein binding site and detecting at least one reporter gene expression.
A kit comprising:
- a) a vertebrate cell with a DNA having a protein binding site, wherein said protein binding site is located at a transcriptionally neutral locus or region of the genome of the vertebrate cell, wherein the transcriptionally neutral locus or region lacks active and repressive histone modifications, and the DNA further comprising at least one reporter gene expression site operatively connected to said protein binding site;
- said kit further comprises b) a DNA vector with a recombinant gene of a binding protein;
- said binding protein being capable of binding to said protein binding site, wherein said binding protein is fused to a member of a Polycomb Repressive Complex (PRC).
A vertebrate cell comprising:
- a DNA having a protein binding site and at least one reporter gene expression site that is operatively connected to said protein binding site;
- wherein said protein binding site is located at a transcriptionally neutral locus or region of the genome of the vertebrate cell, wherein the transcriptionally neutral locus or region lacks active and repressive histone modifications;
- and further comprising a DNA comprising a recombinant gene of a binding protein;
- said binding protein being capable of binding to said protein binding site, wherein said binding protein is fused to a member of a Polycomb Repressive Complex (PRC).
Dependencies are inferred from the claim text.