title: A multiplexed luciferase-based screening platform for interrogating cancer-associated signal transduction in cultured cells.
abstract: Genome-scale interrogation of gene function using RNA interference (RNAi) holds tremendous promise for the rapid identification of chemically tractable cancer cell vulnerabilities. Limiting the potential of this technology is the inability to rapidly delineate the mechanistic basis of phenotypic outcomes and thus inform the development of molecularly targeted therapeutic strategies. We outline here methods to deconstruct cellular phenotypes induced by RNAi-mediated gene targeting using multiplexed reporter systems that allow monitoring of key cancer cell-associated processes. This high-content screening methodology is versatile and can be readily adapted for the screening of other types of large molecular libraries.
authors.affiliation: Department of Cell Biology, UT Southwestern Medical Center.
authors.forename: Ozlem
authors.initials: O
authors.lastname: Kulak
authors.forename: Lawrence
authors.initials: L
authors.lastname: Lum
mesh_terms: D015179:Colorectal Neoplasms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D045325:HCT116 Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D015398:Signal Transduction; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D034741:RNA, Small Interfering; D008156:Luciferases