Article

Cellular arrays for large-scale analysis of transcription factor activity

  1. Abigail D. Bellis1,2,
  2. Beatriz Peňalver-Bernabé1,
  3. Michael S. Weiss1,
  4. Michael E. Yarrington1,
  5. Maria V. Barbolina3,
  6. Angela K. Pannier4,
  7. Jacqueline S. Jeruss5,6,
  8. Linda J. Broadbelt1,
  9. Lonnie D. Shea1,2,6,*

Article first published online: 21 OCT 2010

DOI: 10.1002/bit.22916

Issue

Biotechnology and Bioengineering

Biotechnology and Bioengineering

Volume 108, Issue 2, pages 395–403, February 2011

Additional Information

How to Cite

Bellis, A. D., Peňalver-Bernabé, B., Weiss, M. S., Yarrington, M. E., Barbolina, M. V., Pannier, A. K., Jeruss, J. S., Broadbelt, L. J. and Shea, L. D. (2011), Cellular arrays for large-scale analysis of transcription factor activity. Biotechnol. Bioeng., 108: 395–403. doi: 10.1002/bit.22916

Author Information

  1. 1

    Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd, E156, Evanston, Illinois 60208-3120; telephone: 847-491-7043; fax: 847-491-3728

  2. 2

    Institute for Bionanotechnology in Medicine, Northwestern University, Chicago, Illinois

  3. 3

    Department of Biopharmaceutical Sciences, University of Illinois at Chicago, Chicago, Illinois

  4. 4

    Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska

  5. 5

    Department of Surgery, Northwestern University Feinberg School of Medicine, Chicago, Illinois

  6. 6

    The Robert H. Lurie Comprehensive Cancer Center of Northwestern University, Galter Pavilion, Chicago, Illinois

*Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd, E156, Evanston, Illinois 60208-3120; telephone: 847-491-7043; fax: 847-491-3728.

Publication History

  1. Issue published online: 16 DEC 2010
  2. Article first published online: 21 OCT 2010
  3. Accepted manuscript online: 1 SEP 2010 09:27AM EST
  4. Manuscript Accepted: 17 AUG 2010
  5. Manuscript Revised: 12 JUL 2010
  6. Manuscript Received: 18 APR 2010

Funded by

  • NIH. Grant Numbers: R21 006520, T32 GM008449
  • The Searle Funds at The Chicago Community Trust

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Keywords:

  • cellular array;
  • transcription factor activity;
  • bioluminescence imaging

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. Biotechnol. Bioeng. 2011;108: 395–403. © 2010 Wiley Periodicals, Inc.

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