title: Cellular arrays for large-scale analysis of transcription factor activity.
abstract: Identifying molecular mechanisms or therapeutic targets is typically based on large-scale cellular analysis that measures the abundance of mRNA or protein; however, abundance does not necessarily correlate with activity. We report a method for direct large-scale quantification of active pathways that employs a cellular array with parallel gene delivery of constructs that report pathway activity. The reporter constructs encode luciferase, whose expression is influenced by binding of transcription factors (TFs), which are the downstream targets of signaling pathways. Luciferase levels are quantified by bioluminescence imaging (BLI), which allows for rapid, non-invasive measurements. Activity profiles by BLI of 32 TFs were robust, consistent, and reproducible, and correlated with standard cell lysis techniques. The array identified five TFs with differential activity during phorbol-12-myristate-13-acetate (PMA)-induced differentiation of breast cancer cells. A system for rapid, large-scale, BLI quantification of pathway activity provides an enabling technology for mechanistic studies of cellular responses and processes.
authors.affiliation: Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208-3120, USA.
authors.forename: Abigail D
authors.initials: AD
authors.lastname: Bellis
authors.forename: Beatriz
authors.initials: B
authors.lastname: Peňalver-Bernabé
authors.forename: Michael S
authors.initials: MS
authors.lastname: Weiss
authors.forename: Michael E
authors.initials: ME
authors.lastname: Yarrington
authors.forename: Maria V
authors.initials: MV
authors.lastname: Barbolina
authors.forename: Angela K
authors.initials: AK
authors.lastname: Pannier
authors.forename: Jacqueline S
authors.initials: JS
authors.lastname: Jeruss
authors.forename: Linda J
authors.initials: LJ
authors.lastname: Broadbelt
authors.forename: Lonnie D
authors.initials: LD
authors.lastname: Shea
mesh_terms: D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects*; D017930:Genes, Reporter; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D046228:Microarray Analysis; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D014157:Transcription Factors; D008156:Luciferases
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