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Gold Nanoparticle-Based Förster Resonance Energy Transfer (FRET) Analysis of Estrogen Receptor: DNA Interaction

  • Xiaodi Su
  • Khin Moh Moh Aung
  • Steven Lukman
  • Bin Liu
Part of the Methods in Molecular Biology book series (MIMB, volume 1366)

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.

Key words

Förster resonance energy transfer Gold nanoparticles Water-soluble conjugated polyelectrolytes Transcription factor Protein:DNA interactions Estrogenreceptor 

Notes

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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Xiaodi Su
    • 1
  • Khin Moh Moh Aung
    • 1
  • Steven Lukman
    • 1
  • Bin Liu
    • 1
    • 2
  1. 1.Institute of Materials Research and EngineeringAgency for Science, Technology and Research (A*STAR)InnovisSingapore
  2. 2.Department of Chemical & Biomolecular EngineeringNational University of SingaporeSingaporeSingapore

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