Cellular arrays for large-scale analysis of transcription factor activity

Abigail D. Bellis, Beatriz Peňalver-Bernabé, Michael S. Weiss, Michael E. Yarrington, Maria V. Barbolina, Angela K. Pannier, Jacqueline S. Jeruss, Linda J. Broadbelt, Lonnie D. Shea

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)395-403
Number of pages9
JournalBiotechnology and Bioengineering
Volume108
Issue number2
DOIs
StatePublished - Feb 2011

Keywords

  • Bioluminescence imaging
  • Cellular array
  • Transcription factor activity

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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