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- Title
A genome-wide functional assay of signal transduction in living mammalian cells.
- Authors
Whitney, Mike; Rockenstein, Edward; Cantin, Greg; Knapp, Tom; Zlokarnik, Gregor; Sanders, Pam; Durick, Kyle; Craig, Frank F.; Negulescu, Paul A.
- Abstract
We describe a genome-wide functional assay for rapid isolation of cell clones and genetic elements responsive to specific stimuli. A promoterless β-lactamase reporter gene was transfected into a human T-cell line to generate a living library of reporter-tagged clones. When loaded with a cell-permeable fluo-rogenic substrate, the cell library simultaneously reports the expression of a large number of endogenous genes. Flow cytometry was used to recover individual clones whose reporter-tagged genes were either induced or repressed following T-cell activation. Responsive clones were expanded and analyzed pharmacologically to identify patterns of regulation associated with specific genes. Although demonstrated using T cells, the genomic assay could be applied to map downstream transcriptional consequences for any propagating cell line in response to any stimulus of interest.
- Subjects
T cells; BETA lactamases; FLOW cytometry
- Publication
Nature Biotechnology, 1998, Vol 16, Issue 13, p1329
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/4302