title: A genome scale overexpression screen to reveal drug activity in human cells.
abstract: Target identification is a critical step in the lengthy and expensive process of drug development. Here, we describe a genome-wide screening platform that uses systematic overexpression of pooled human ORFs to understand drug mode-of-action and resistance mechanisms. We first calibrated our screen with the well-characterized drug methotrexate. We then identified new genes involved in the bioactivity of diverse drugs including antineoplastic agents and biologically active molecules. Finally, we focused on the transcription factor RHOXF2 whose overexpression conferred resistance to DNA damaging agents. This approach represents an orthogonal method for functional screening and, to our knowledge, has never been reported before.
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Banting and Best Department of Medical Research, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Anthony
authors.initials: A
authors.lastname: Arnoldo
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Banting and Best Department of Medical Research, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Saranya
authors.initials: S
authors.lastname: Kittanakom
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Banting and Best Department of Medical Research, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada ; Donnelly Sequencing Center, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Lawrence E
authors.initials: LE
authors.lastname: Heisler
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Anthony B
authors.initials: AB
authors.lastname: Mak
authors.affiliation: Institute of Biomaterials and Biomedical Engineering, University of Toronto, 170 College Street, Toronto M5S 3E3, Canada.
authors.forename: Andrey I
authors.initials: AI
authors.lastname: Shukalyuk
authors.affiliation: Donnelly Sequencing Center, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Dax
authors.initials: D
authors.lastname: Torti
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada.
authors.forename: Jason
authors.initials: J
authors.lastname: Moffat
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada ; Department of Pharmaceutical Sciences, University of Toronto, 144 College Street, Toronto, Ontario M5S 3M2, Canada ; Department of Pharmaceutical Sciences, University of British Columbia, 6619-2405 Wesbrook Mall, Vancouver, BC V6T 1Z3, Canada.
authors.forename: Guri
authors.initials: G
authors.lastname: Giaever
authors.affiliation: Department of Molecular Genetics, University of Toronto, Toronto, M5S 3E1, Canada ; Banting and Best Department of Medical Research, University of Toronto, Toronto, M5S 3E1, Canada ; Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada ; Donnelly Sequencing Center, University of Toronto, 160 College Street, Toronto, Ontario M5S 3E1, Canada ; Department of Pharmaceutical Sciences, University of British Columbia, 6619-2405 Wesbrook Mall, Vancouver, BC V6T 1Z3, Canada.
authors.forename: Corey
authors.initials: C
authors.lastname: Nislow