Methods for Screening for Compounds that Modulate Insulin Promoter Activity
AssigneeBurnham Institute for Medical Research
InventorsMark Mercola, Fred Levine
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Abstract
Compositions and methods are provided for screening for compounds that modulate insulin promoter activity. Vectors that express green fluorescent protein under the control of the human insulin promoter are introduced into mouse and human cells in which the insulin promoter is expressed in a glucose-responsive manner. Such cells are then used to screen for compounds that modulate insulin promoter activity.
Claims45
A vector comprising a human insulin gene promoter polynucleotide sequence operably linked to a polynucleotide that encodes a destabilized enhanced green fluorescent protein polynucleotide sequence, wherein destabilized enhanced green fluorescent protein is expressed upon introduction of the vector into a cell selected from the group consisting of a MIN6 mouse insulinoma cell and a human T6PN/E47MER cell.
Lentiviral vector pRRL.SIN-18.cPPT.hINS-EGFP.WPRE.
A human T6PN cell comprising a polynucleotide sequence that expresses E47.
A human T6PN/E47MER cell.
A method of identifying a compound that modulates insulin gene expression, the method comprising:
- (a) providing a cell comprising a vector that comprises a human insulin gene promoter polynucleotide that is operably linked to an enhanced green fluorescent protein polynucleotide, wherein enhanced green fluorescent protein is expressed at a baseline level in the cell;
- (b) contacting the cell with a candidate compound;
- and (c) detecting a modulation in expression of enhanced green fluorescent protein in the cell compared to the baseline level as a result of contacting the cell with the candidate compound.
A method of identifying a compound that modulates insulin gene expression, the method comprising:
- (a) providing a cell selected from the group consisting of a MIN6 mouse insulinoma cell and a human T6PN/E47MER cell, the cell comprising a vector that comprises a human insulin gene promoter polynucleotide that is operably linked to an enhanced green fluorescent protein polynucleotide, wherein enhanced green fluorescent protein is expressed at a baseline level in the cell;
- (b) contacting the cell with a candidate compound;
- and (c) detecting a modulation in expression of enhanced green fluorescent protein in the cell compared to the baseline level as a result of contacting the cell with the candidate compound.
Dependencies are inferred from the claim text.