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Updated: May 30, 2026

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy
Published on: February 7, 2019
Binding force measurement of NF-κB-ODNs interaction: an AFM based decoy and drug testing tool
Michele Menotta1, Rita Crinelli, Elisa Carloni
1Department of Biomolecular Sciences, University of Urbino Carlo Bo, Via Saffi 2, 61029 Urbino (PU), Italy.
Abstract:
Interaction between transcription factors and DNA are essential for regulating gene transcription. The Nuclear factor-κB (NF-κB) is a ubiquitous transcription factor involved in cell signalling and its failure is a principal cause of several autoimmune and auto-inflammatory disorders. In this paper we have developed an atomic force microscopy (AFM) method to quantitatively characterise the interaction force between NF-κB and DNA or LNA (locked nucleic acid) double strand molecules containing the NF responsive elements (RE). This process allows the simple testing and selection of LNA based decoy molecules to be used in NF-κB modulation decoy strategies. Furthermore the proposed methodology is also suitable for testing drug efficacy on the modulation of NF-κB binding to its nucleic acid target sequence. A biological AFM based sensor is therefore considered appropriate for characterising transcription factors and selecting molecules to modulate their activity.
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