QUANTITATION AND ARCHIVING OF GENE EXPRESSION

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Tumor malignant cell populations frequently occupy less than 5% of the tissue volume. Laser capture microdissection (LCM) provides a means of selecting malignant cells of similar morphology, but highly sensitive and accurate methods are needed for measuring DNA content, protein and mRNA expression levels ideally all within the same small number of microdissected cells. Molecular Innovations, incorporated (MII) will develop a precise method for simultaneous measurement of gene amplification and expression from LCM cells. Precise gene measurement is accomplished through the design of specific amplification primers and probes that provide a means of normalizing measurement fluctuations due to cell number, pseudogene sequences, specimen fixation, nucleic acid extraction, RNA degradation, and PCR amplification efficiency. Extracted Nucleic Acids are bound irreversibly to a solid phase material and can be repeatedly analyzed in series for multiple genes (archiving). The first gene targets include Estrogen and Progesterone Receptors (ER and PgR) and the c-erbB-2 oncogene. These genes are well accepted as important in the clinical management of breast cancer. Our assay will provide for more precise gene expression measurement localized in the specific tissue cells relevant to the breast tumor pathology. PROPOSED COMMERCIAL APPLICATIONS: Breast cancer is a leading cause of cancer deaths in women. An improved and more precise method of detecting gene expression levels that have been shown to be of prognostic and therapeutic importance has great commercialization potential.