Methods and compositions for screening for altered cellular phenotypes

Patent Number: 7056687

This patent is useful for drug development. It teaches ways of inducing gene expression in cells (including stem cells) and using the specifically modified cells for screening of drug libraries. The invention itself is not specific to stem cells, so therefore we will not describe it in detail. The patent has 3 independent claims. Specifically:

"A method of screening for cells having an altered phenotype, the method comprising the steps of: a) providing a population of cells having a parent phenotype, said population of cells comprising a nucleic acid sequence encoding a first element expressed in said population of cells; b) introducing into said population of cells a library of fusion nucleic acids, said fusion nucleic acids each comprising: (1) a second element that is regulatable by said first element; and (2) a nucleic acid sequence encoding a candidate bioactive agent, wherein said nucleic acid sequence is operably linked to said second element; c) modulating the expression of said nucleic acid sequence encoding the candidate bioactive agent by contacting said population of cells with a third element, wherein said modulation is selected from the group consisting of induction and repression of said nucleic acid sequence encoding the candidate bioactive agent; d) collecting a first subpopulation of cells having an altered phenotype; e) modulating in the opposite direction of step © the expression of said nucleic acid sequence by modulating said contacting of first subpopulation of cells with said third element; f) collecting a second subpopulation of cells having said parent phenotype; g) modulating in the same direction as in step © the expression of said nucleic acid sequence by contacting said second subpopulation of cells with said third element; and h) detecting a third subpopulation of cells having said altered phenotype."

"A method of screening for cells having an altered phenotype, the method comprising the steps of: a) providing a population of cells having a parent phenotype, said population of cells comprising a nucleic acid sequence encoding a first element expressed in said population of cells; b) introducing into said population of cells a library of fusion nucleic acids, said fusion nucleic acids each comprising: 1) a second element that is regulatable by said first element; and 2) a nucleic acid sequence encoding a candidate bioactive agent, wherein said nucleic acid sequence is operably linked to said second element; c) inducing the expression of said nucleic acid sequence encoding the candidate bioactive agent by contacting said population of cells with a third element; d) collecting a first subpopulation of cells having an altered phenotype; e) repressing the expression of said nucleic acid sequence by modulating said contacting of first subpopulation of cells with said third element; f) collecting a second subpopulation of cells having said parent phenotype; g) inducing the expression of said nucleic acid sequence by contacting said second subpopulation of cells with said third element; and h) detecting a third subpopulation of cells having said altered phenotype. "

"A method of screening for cells having an altered phenotype, the method comprising the steps of: a) providing a population of cells having a parent phenotype, said population of cells comprising a nucleic acid sequence encoding a first element; b) introducing into said population of cells a library of fusion nucleic acids, said fusion nucleic acids each comprising: (1) a second element that is regulatable by said first element; and (2) a nucleic acid sequence encoding a candidate bioactive agent, wherein said nucleic acid sequence is operably linked to said second element; c) inducing the expression of said nucleic acid sequence encoding the candidate bioactive agent by expressing said first element in said population of cells; d) collecting a first subpopulation of cells having an altered phenotype; e) repressing the expression of said nucleic acid sequence by contacting said first subpopulation of cells with a third element; f) collecting a second subpopulation of cells having said parent phenotype; g)inducing the expression of said nucleic acid sequence by modulating said contacting of said second subpopulation of cells with said third element; and h) detecting a third subpopulation of cells having said altered phenotype. "

View this patent on the USPTO website.