Abstract
Artificial transcription factors (ATFs) consist of a transcriptional effector domain fused to a DNA-binding domain such as an engineered zinc finger protein (ZFP). Depending on the effector domain, ATFs can up- or downregulate gene expression and thus represent powerful tools in biomedical research and allow novel approaches in clinical practice. Here, we describe the construction of ATFs directed against the promoter of the epithelial cell adhesion molecule and against the promoter of the RNA component of telomerase. Methods to assess DNA binding of the engineered ZFP as well as to determine and improve the cellular effect of ATFs on (endogenous) promoter activity are described.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Visser, A.E., Verschure, P.J., Gommans, W.M., Haisma, H.J., and Rots, M.G. (2006) Step into the groove: engineered transcription factors as modulators of gene expression. Adv Genet. 56, 131–161.
Lu, Y., Tian, C., Danialou, G., Gilbert, R., Petrof, B.J., Karpati, G., and Nalbantoglu, J. (2008) Targeting artificial transcription factors to the utrophin A promoter: effects on dystrophic pathology and muscle function. J Biol Chem. 283, 34720–34727.
Beltran, A., Parikh, S., Liu, Y., Cuevas, B.D., Johnson, G.L., Futscher, B.W., and Blancafort, P. (2007) Re-activation of a dormant tumor suppressor gene maspin by designed transcription factors. Oncogene. 26, 2791–2798.
Kang, Y.A., Shin, H.C., Yoo, J.Y., Kim, J.H., Kim, J.S., and Yun, C.O. (2008) Novel cancer antiangiotherapy using the VEGF promoter-targeted artificial zinc-finger protein and oncolytic adenovirus. Mol Ther. 16, 1033–1040.
Sera, T. (2009) Zinc-finger-based artificial transcription factors and their applications. Adv Drug Deliv Rev. 61, 513–526.
Verschure, P.J., Visser, A.E., and Rots, M.G. (2006) Step out of the groove: epigenetic gene control systems and engineered transcription factors. Adv Genet. 56, 163–204.
Beltran, A.S., Sun, X., Lizardi, P.M., and Blancafort, P. (2008) Reprogramming epigenetic silencing: artificial transcription factors synergize with chromatin remodeling drugs to reactivate the tumor suppressor mammary serine protease inhibitor. Mol Cancer Ther. 7, 1080–1090.
Carroll, D., Morton, J.J., Beumer, K.J., and Segal, D.J. (2006) Design, construction and in vitro testing of zinc finger nucleases. Nat Protoc. 1, 1329–1341.
Gommans, W.M., McLaughlin, P.M., Lindhout, B.I., Segal, D.J., Wiegman, D.J., Haisma, H.J., van der Zaal, B.J., and Rots, M.G. (2007) Engineering zinc finger protein transcription factors to downregulate the epithelial glycoprotein-2 promoter as a novel anti-cancer treatment. Mol Carcinog. 46, 391–401.
Zhao, J.Q., Hoare, S.F., McFarlane, R., Muir, S., Parkinson, E.K., Black, D.M., and Keith, W.N. (1998) Cloning and characterization of human and mouse telomerase RNA gene promoter sequences. Oncogene. 16, 1345–1350.
Shay, J.W. and Keith, W.N. (2008) Targeting telomerase for cancer therapeutics. Br J Cancer. 98, 677–683.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Stolzenburg, S., Bilsland, A., Keith, W.N., Rots, M.G. (2010). Modulation of Gene Expression Using Zinc Finger-Based Artificial Transcription Factors. In: Mackay, J., Segal, D. (eds) Engineered Zinc Finger Proteins. Methods in Molecular Biology, vol 649. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-753-2_7
Download citation
DOI: https://doi.org/10.1007/978-1-60761-753-2_7
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60761-752-5
Online ISBN: 978-1-60761-753-2
eBook Packages: Springer Protocols