Patent ID: 6197521
Filing Date: 2001-03-06
Classification: A61K,A61P,C07K,C12N,C12Y,G01N

Abstract:
A method for performing an assay comprising the steps of:(a) isolating cellular RNA comprising messenger RNA from cancer cells from an animal;(b) measuring an expression level of an mRNA that encodes a gene that is homologous to a nucleotide sequence of a genetic suppressor element (GSE) or its complement in the cancer cells from the animal, wherein the genetic suppressor element is a genetic suppressor element associated with establishment or maintenance of a transformed phenotype in a mammalian cell, wherein the genetic suppressor element is produced by a method comprising the steps of:(i) synthesizing randomly fragmented cDNA prepared from the total mRNA of a cell to yield DNA fragments;(ii) transferring the DNA fragments to an expression vector to yield a genetic suppressor element library, wherein the expression vector is capable of expressing the DNA fragments in a living cell that is capable of expressing a transformed phenotype;(iii) genetically modifying the living cells by introducing the genetic suppressor element library into the cells;(iv) isolating or enriching for genetically modified living cells containing genetic suppressor elements conferring the transformed phenotype on the cells by selecting the cells under conditions wherein the transformed cells are identifiable; and(v) obtaining the genetic suppressor element conferring the transformed phenotype from the surviving genetically modified cells;wherein the assay further comprises the step of:(c) determining whether the level of expression of said mRNA measured in subpart (b) indicates that the gene is over-expressed or under-expressed in the cancer cells of the animal.