Assay For Transcriptionally Active Methylated Promoters
AssigneeUniversity of South Florida
InventorsGeorge Blanck, Melissa Niesen, Aaron Osborne
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Abstract
A method of identifying genes that can be activated when methylated; as well as proteins that mediate such activation. The method allows identification of HLA-DRA silencing agents expected to be of therapeutic value in the treatment of inflammation associated with HLA-DRA expression.
Claims20
A method of identifying a transcriptional activation mediator of a methylated gene of interest, comprising the steps of:
- providing a cell comprising a methylated construct further comprising a reporter-gene operably linked to a promoter of the gene of interest;
- administering a test agent to the cell;
- and
- comparing the expression of the reporter gene to a control to determine the test agent's ability to mediate transcriptional activation of the methylated gene.
A method of identifying a gene of interest capable of methylation-dependent activation, comprising the steps of;
- providing a cell comprising the gene of interest;
- establishing the gene of interest is transcriptionally inactive responsive to DNA methylization associated therewith;
- administering a methylated expression construct to the cell, comprising a promoter of the gene of interest operably linked to a reporter gene;
- determining the expression of the gene of the interest, compared to a control, wherein a comparative increase in the expression of the gene of interest in the cell indicates methylation-dependent activation thereof.
A method of screening for an agent as a candidate for treatment of inflammation associated with the expression of a methylated gene of interest, comprising the steps of:
- providing a cell comprising a methylated construct further comprising a reporter-gene operably linked to a promoter of the gene of interest;
- transfecting the cell with a DNA binding protein specific to the gene of interest;
- contacting the cell with a test agent;
- and
- measuring the expression of the reporter gene compared to a control as an indicator of the test agent's ability to modulate expression of the gene of interest.
Dependencies are inferred from the claim text.