Methods of determining chemicals that modulate transcriptionally expression of genes associated with cardiovascular disease

Claim 1

A method of determining whether a chemical not previously known to be a modulator of protein biosynthesis is capable of specifically transcriptionally modulating the expression of a gene encoding a protein of interest associated with the treatment of one or more symptoms of a cardiovascular disease which comprises: (a) contacting a sample which contains a predefined number of identical eucaryotic cells with a predetermined concentration of the chemical to be tested, each such cell comprising a single DNA construct consisting essentially of in 5' to 3' order (i) a modulatable transcriptional regulatory sequence of the gene encoding the protein of interest, (ii) a promoter of the gene encoding the protein of interest, and (iii) a reporter gene which expresses a polypeptide capable of producing a detectable signal, coupled to, and under the control of, the promoter, under conditions such that the chemical if capable of acting as a transcriptional modulator of the gene encoding the protein of interest, causes a measurable detectable signal to be produced by the polypeptide expressed by the reporter gene; (b) quantitatively determining the amount of the signal so produced; and (c) comparing the amount so determined with the amount of produced signal detected in the absence of any chemical being tested and upon contacting the sample with other chemicals so as to thereby identify the chemical as a chemical which causes a change in the detectable signal produced by the polypeptide, and determining whether the chemical specifically transcriptionally modulates expression of the gene associated with the treatment of one or more symptoms of the cardiovascular disease.

Claim 2

A method of determining whether a chemical not previously known to be a modulator of protein biosynthesis is capable of specifically transcriptionally modulating the expression of a gene encoding a protein of interest associated with the treatment of one or more symptoms of a cardiovascular disease which comprises: (a) contacting a sample which contains a predefined number of identical eucaryotic cells with a predetermined concentration of the chemical to be tested, each such cell comprising a single DNA construct consisting essentially of in 5' to 3' order (i) a modulatable transcriptional regulatory sequence of the gene encoding the protein of interest, (ii) a promoter of the gene encoding the protein of interest, and (iii) a reporter gene which expresses a polypeptide capable of producing a detectable signal, coupled to, and under the control of, the promoter, under conditions such that the chemical if capable of acting as a transcriptional modulator of the gene encoding the protein of interest, causes a measurable detectable signal to be produced by the polypeptide expressed by the reporter gene; (b) quantitatively determining the amount of the signal so produced; and (c) comparing the amount so determined with the amount of produced signal detected in the absence of any chemical being tested and upon contacting the sample with other chemicals so as to thereby identify the chemical as a chemical which causes a change in the detectable signal produced by the polypeptide, and determining whether the chemical specifically transcriptionally modulates expression of the gene associated with the treatment of one or more symptoms of the cardiovascular disease.

Claim 3

A method of determining whether a chemical not previously known to be a modulator of protein biosynthesis is capable of specifically transcriptionally modulating the expression of a gene encoding a protein of interest associated with the treatment of one or more symptoms of a cardiovascular disease which comprises: (a) contacting a sample which contains a predefined number of identical eucaryotic cells with a predetermined concentration of the chemical to be tested, each such cell comprising a single DNA construct consisting essentially of in 5' to 3' order (i) a modulatable transcriptional regulatory sequence of the gene encoding the protein of interest, (ii ) a promoter of the gene encoding the protein of interest, and (iii) a DNA sequence transcribable into mRNA coupled to, and under the control of, the promoter, under conditions such that the chemical if capable of acting as a transcriptional modulator of the gene encoding the protein of interest, causes a measurable difference in the amount of mRNA transcribed from the DNA sequence; (b) quantitatively determining the amount the mRNA so produced; and (c) comparing the amount so determined with the amount of mRNA detected in the absence of any chemical being tested and upon contacting the sample with other chemicals so as to thereby identify the chemical as a chemical which causes a change in the detectable mRNA amount, and determining whether the chemical specifically transcriptionally modulates expression of the gene at interest associated with the treatment of one or more symptoms of the cardiovascular disease.

Claim 4

A method of determining whether a chemical not previously known to be a modulator of protein biosynthesis is capable of specifically transcriptionally modulating the expression of a gene encoding a protein of interest associated with the treatment of one or more symptoms of a cardiovascular disease which comprises: (a) contacting a sample which contains a predefined number of identical eucaryotic cells with a predetermined concentration of the chemical to be tested, each such cell comprising a single DNA construct consisting essentially of in 5' to 3' order (i) a modulatable transcriptional regulatory sequence of the gene encoding the protein of interest, (ii ) a promoter of the gene encoding the protein of interest, and (iii) a DNA sequence transcribable into mRNA coupled to, and under the control of, the promoter, under conditions such that the chemical if capable of acting as a transcriptional modulator of the gene encoding the protein of interest, causes a measurable difference in the amount of mRNA transcribed from the DNA sequence; (b) quantitatively determining the amount the mRNA so produced; and (c) comparing the amount so determined with the amount of mRNA detected in the absence of any chemical being tested and upon contacting the sample with other chemicals so as to thereby identify the chemical as a chemical which causes a change in the detectable mRNA amount, and determining whether the chemical specifically transcriptionally modulates expression of the gene at interest associated with the treatment of one or more symptoms of the cardiovascular disease.