Dna constructs and methods for screening for increased expression of

Abstract

Methods for screening for a drug that increases expression of the human apolioprotein (apo) AI gene and related DNA constructs. A first method has the steps of: (a) introducing into mammalian cells a DNA construct including, functionally joined together in the 5'.fwdarw.3' direction of transcription, (i) at least one copy of a drug-responsive element (DRE) having at least about 80% homology with the DNA sequence 5'-G/C N T/G A/G GCTGGG-3', (ii) a heterologous promoter, (iii) a reporter gene and (iv) an untranslated region including a functional polyadenylation signal; (b) growing a first culture of the cells in the absence of drug; (c) lysing the first culture to produce a first extract; (d) assaying the first extract for activity of a protein encoded by the reporter gene; (e) growing a second culture of the cells in the presence of the drug; (f) lysing the second culture to produce a second extract; (g) assaying the second extract for activity of the protein encoded by the reporter gene; and (h) comparing the activities of the first extract and the second extract. A second method uses a different DNA construct including (i) a promoter region of the human apo AI gene including at least one copy of a drug-responsive element (DRE) having at least about 80% homology with the DNA sequence 5'-G/C N T/G A/G GCTGGG-3', (ii) a reporter gene and (iii) an untranslated region including a functional polyadenylation signal. A third method involves stably maintaining the introduced DNA in the cells.