Direct quantification of gene expression using capillary electrophoresis with laser-induced fluorescence - Patents

Direct quantification of gene expression using capillary electrophoresis with laser-induced fluorescence

Agent: Needle & Rosenberg, P.C. - Atlanta, GA, US
Inventors: Edie C. Goldsmith, Jack G. Goldsmith
Applicaton #: 20060286552
Class: 435005000 (USPTO)
12/21/06
Provided is a method for direct quantification of gene expression using capillary electrophoresis with laser-induced fluorescence to measure RNA in a sample. Also provided is a method of diagnosing a disease in a subject, wherein the disease is caused by increased or decreased expression of a causative gene.
BACKGROUND OF THE INVENTION
[0003] 1. Field of the Invention
[0004] This invention relates generally to the field of measuring gene expression. Specifically, the present invention relates to a novel method for directly quantifying gene expression by measuring RNA using capillary electrophoresis with laser-induced fluorescence.
SUMMARY OF THE INVENTION
[0009] Capillary electrophoresis with laser induced fluorescence (CE-LIF) is a novel, method for the direct quantitation of gene expression. CE-LIF provides a fast, direct, sensitive and non-radioactive means to detect molecules, for example RNA. The method described herein is applicable to any RNA for which some sequence information is known and represents the first direct quantitation of gene expression in which the size of the target gene is obtained.
[0010] In accordance with the purposes of this invention, as embodied and broadly described herein, this invention, in one aspect, relates to a method of measuring an amount of ribonucleic acid (RNA), comprising (a) incubating an RNA sample with a fluorescently labeled RNA probe complementary to a sequence in a target RNA under conditions whereby the probe can hybridize to the target RNA to form an RNA:fluorescently labeled probe hybrid, (b) passing the RNA:fluorescently labeled probe hybrid through a capillary electrophoresis system, (c) detecting and recording changes in fluorescence as a peak as the RNA:fluorescently labeled probe passes through a detection window, and (d) determining an area under the peak, whereby the area under the peak indicates the amount of target RNA.
[0011] In another aspect, the invention relates to a method of diagnosing a disease in a subject, wherein the disease is characterized by an increased or decreased level of expression of a gene, comprising (a) determining a level of expression of the gene in the subject according to the disclosed method; (b) comparing the level of expression of the gene in the subject to a control level of expression of a gene from a subject without disease; (c) determining whether the level of expression from the subject in step (a) is increased or decreased compared to the control level, whereby an increased or decreased level of expression of the gene in the subject diagnoses the disease in the subject.
[0012] In another aspect, the invention relates to a method of diagnosing a bacterial disease or a viral disease in a subject, comprising identifying a target bacterial RNA or a target viral RNA in a sample from the subject according to the disclosed method, wherein the identification of the target bacterial RNA or the target viral RNA diagnoses the bacterial disease or the viral disease in the subject.
[0013] Additional advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The advantages of the invention will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.
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