Bioluminescence imaging for assessment and normalization in transfected cell arrays

Angela K. Pannier, Eric A. Ariazi, Abigail D. Bellis, Zain Bengali, V. Craig Jordan, Lonnie D. Shea

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Transfected cell arrays (TCAs) represent a high-throughput technique to correlate gene expression with functional cell responses. Despite advances in TCAs, improvements are needed for the widespread application of this technology. We have developed a TCA that combines a two-plasmid system and dual-biolumineseence imaging to quantitatively normalize for variability in transfection and increase sensitivity. The two-plasmids consist of: (i) normalization plasmid present within each spot, and (ii) functional plasmid that varies between spots, responsible for the functional endpoint of the array. Bioluminescence imaging of dual-luciferase reporters (renilla, firefly luciferase) provides sensitive and quantitative detection of cellular response, with minimal post-transfection processing. The array was applied to quantify estrogen receptor α (ERα) activity in MCF-7 breast cancer cells. A plasmid containing an ERα-regulated promoter directing firefly luciferase expression was mixed with a normalization plasmid, compfexed with cationic lipids and deposited into an array. ER induction mimicked results obtained through traditional assays methods, with estrogen inducing luciferase expression 10-fold over the antiestrogen fulvestrant or vehicle. Furthermore, the array captured a dose response to estrogen, demonstrating the sensitivity of bioluminescence quantification. This system provides a tool for basic science research, with potential application for the development of patient specific therapies.

Original languageEnglish (US)
Pages (from-to)486-497
Number of pages12
JournalBiotechnology and Bioengineering
Volume98
Issue number2
DOIs
StatePublished - Oct 1 2007

Fingerprint

Bioluminescence
Plasmids
Imaging techniques
Estrogen Receptors
Firefly Luciferases
Luciferases
Transfection
Estrogens
Renilla Luciferases
Estrogen Receptor Modulators
Gene expression
Lipids
Assays
Cells
Throughput
Breast Neoplasms
Technology
Gene Expression
Processing
Research

Keywords

  • Bioluminescence imaging
  • Breast cancer
  • Estrogen receptor
  • Substrate-mediated gene delivery
  • Transfected cell array

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Bioluminescence imaging for assessment and normalization in transfected cell arrays. / Pannier, Angela K.; Ariazi, Eric A.; Bellis, Abigail D.; Bengali, Zain; Jordan, V. Craig; Shea, Lonnie D.

In: Biotechnology and Bioengineering, Vol. 98, No. 2, 01.10.2007, p. 486-497.

Research output: Contribution to journalArticle

Pannier, Angela K. ; Ariazi, Eric A. ; Bellis, Abigail D. ; Bengali, Zain ; Jordan, V. Craig ; Shea, Lonnie D. / Bioluminescence imaging for assessment and normalization in transfected cell arrays. In: Biotechnology and Bioengineering. 2007 ; Vol. 98, No. 2. pp. 486-497.
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