Abstract
Estrogen receptors play critical roles in regulating genes responsible for development and maintenance of reproductive tissues and other physiological function. The interaction of ERs with DNA sequences, known as estrogen response elements (EREs) (a palindromic repeat separated by three-base spacer, 5′GGTCAnnnTGACC-3′), is required for estrogen regulation of target gene expression. Here, we describe a simple “mix-and-measure”-based method for detecting ER:ERE interactions using ERE-immobilized metal nanoparticles and water-soluble conjugated polyelectrolytes (CPEs) as cooperative sensing elements. This method can differentiate the distinct DNA-binding affinity between ERα and ERβ, and determine ER:ERE-binding stoichiometry. This method can also accurately detect all 15 singly mutated EREs (i.e., three possible base substitutions at each of one to five positions from left to right of the 5′ end half site, GGTCA) for their binding energy to ER. This method is compatible with 96-well plate format for high-throughput analysis.
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Acknowledgements
The authors would like to acknowledge the Agency for Science, Technology and Research (A*STAR), Singapore, for the financial support (JCO 1131CFG001).
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Su, X., Aung, K.M.M., Lukman, S., Liu, B. (2016). Gold Nanoparticle-Based Förster Resonance Energy Transfer (FRET) Analysis of Estrogen Receptor: DNA Interaction. In: Eyster, K.M. (eds) Estrogen Receptors. Methods in Molecular Biology, vol 1366. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3127-9_17
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DOI: https://doi.org/10.1007/978-1-4939-3127-9_17
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3126-2
Online ISBN: 978-1-4939-3127-9
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