| 摘要 |
A method for the identification of new therapeutic targets and protein interaction sites for use in drug discovery. In particular the invention relates to a method for identifying inhibitors of a cell signalling pathway, the method comprising (1) providing a population of mammalian cells, each mammalian cell having an active cell signalling pathway and comprising: (a) a first heterologous nucleic acid comprising; (i) a nucleotide sequence encoding a first detectable reporter and, (ii) a constitutive regulatory element which is operably linked to the nucleotide sequence; and, (b) a second heterologous nucleic acid comprising: (i) a first nucleotide sequence encoding a repressor molecule, for example an RNA or protein, which inactivates, inhibits or suppresses expression of the first detectable reporter, (ii) a second nucleotide sequence encoding a second detectable reporter; and (iii) a signal-activated regulatory element which is activated by said cell signalling pathway, said signal-activated regulatory element being operably linked to the first and second nucleotide sequences, (2) introducing a library of test compounds into said population of mammalian cells, and; (3) determining the expression of the first and the second detectable reporters in one or more of the population of transfected cells, wherein expression of the first detectable reporter but not the second detectable reporter in a transfected cell is indicative that the test biomolecule expressed by the nucleic acid in the cell inhibits said cell signalling pathway. |
