Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment
✍ Scribed by Willemijn M. Gommans; Pamela M.J. McLaughlin; Beatrice I. Lindhout; David J. Segal; D.J. Wiegman; Hidde J. Haisma; Bert J. van der Zaal; Marianne G. Rots
- Publisher
- John Wiley and Sons
- Year
- 2007
- Tongue
- English
- Weight
- 239 KB
- Volume
- 46
- Category
- Article
- ISSN
- 0899-1987
- DOI
- 10.1002/mc.20289
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✦ Synopsis
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. © 2006 Wiley‐Liss, Inc.