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

20 Citations (Scopus)

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

Fingerprint

Zinc Fingers
Glycoproteins
Transcription Factors
Down-Regulation
Neoplasms
Proteins
Nucleotide Motifs
Gene Expression
Technology
DNA

Keywords

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

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology

Cite this

Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment. / Gommans, Willemijn M.; McLaughlin, Pamela M J; Lindhout, Beatrice I.; Segal, David; Wiegman, D. J.; Haisma, Hidde J.; Van Der Zaal, Bert J.; Rots, Marianne G.

In: Molecular Carcinogenesis, Vol. 46, No. 5, 05.2007, p. 391-401.

Research output: Contribution to journalArticle

Gommans, WM, McLaughlin, PMJ, Lindhout, BI, Segal, D, Wiegman, DJ, Haisma, HJ, Van Der Zaal, BJ & Rots, MG 2007, 'Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment', Molecular Carcinogenesis, vol. 46, no. 5, pp. 391-401. https://doi.org/10.1002/mc.20289
Gommans, Willemijn M. ; McLaughlin, Pamela M J ; Lindhout, Beatrice I. ; Segal, David ; Wiegman, D. J. ; Haisma, Hidde J. ; Van Der Zaal, Bert J. ; Rots, Marianne G. / Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment. In: Molecular Carcinogenesis. 2007 ; Vol. 46, No. 5. pp. 391-401.
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