Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment

Willemijn M. Gommans, Pamela M J McLaughlin, Beatrice I. Lindhout, David Segal, D. J. Wiegman, Hidde J. Haisma, Bert J. Van Der Zaal, Marianne G. Rots

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Zinc finger protein transcription factors (ZFP-TFs) are emerging as powerful novel tools for the treatment of many different diseases. ZFPs are DNA-binding motifs and consist of modular zinc finger domains. Each domain can be engineered to recognize a specific DNA triplet, and stitching six domains together results in the recognition of a gene-specific sequence. Inhibition of gene expression can be achieved by fusing a repressor domain to these DNA-binding motifs. In this study, we engineered ZFP-TFs to downregulate the activity of the epithelial glycoprotein-2 (EGP-2) promoter. The protein EGP-2 is overexpressed in a wide variety of cancer types and EGP-2 downregulation has been shown to result in a decreased oncogenic potential of tumor cells. Therefore, downregulation of EGP-2 expression by ZFP-TFs provides a novel anti-cancer therapeutic. Using a straightforward strategy, we engineered a 3-ZFP that could bind a 9 bp sequence within the EGP-2 promoter. After the addition of a repressor domain, this 3-ZFP-TF could efficiently downregulate EGP-2 promoter activity by 60%. To demonstrate the flexibility of this technology, we coupled an activation domain to the engineered ZFP, resulting in a nearly 200% increase in EGP-2 promoter activity. To inhibit the endogenous EGP-2 promoter, we engineered 6-ZFP-TFs. Although none of the constructed ZFP-TFs could convincingly modulate the endogenous promoter, efficient and specific inhibition of the exogenous promoter was observed. Overall, ZFP-TFs are versatile bi-directional modulators of gene expression and downregulation of EGP-2 promoter activity using ZFP-TFs can ultimately result in a novel anti-cancer treatment.

Original languageEnglish (US)
Pages (from-to)391-401
Number of pages11
JournalMolecular Carcinogenesis
Volume46
Issue number5
DOIs
StatePublished - May 2007

Keywords

  • Artificial transcription factors
  • EpCAM
  • Gene expression regulation
  • Silencing

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology

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    Gommans, W. M., McLaughlin, P. M. J., Lindhout, B. I., Segal, D., Wiegman, D. J., Haisma, H. J., Van Der Zaal, B. J., & Rots, M. G. (2007). Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment. Molecular Carcinogenesis, 46(5), 391-401. https://doi.org/10.1002/mc.20289