Single-Molecule DNA Biosensors for Protein and Ligand Detection

Authors

  • Konstantinos Lymperopoulos Dr.,

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
    2. Current address: BioQuant Institute, Cellnetworks Cluster, Ruprecht-Karls Universität Heidelberg, 69120 Heidelberg (Germany)
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    • These authors contributed equally to this work.

  • Robert Crawford,

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
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    • These authors contributed equally to this work.

  • Joseph P. Torella,

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
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  • Mike Heilemann Dr.,

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
    2. Current address: Applied Laser Physics and Laser Spectroscopy, Bielefeld University, Universitätsstrasse 25, 33615 Bielefeld (Germany)
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  • Ling Chin Hwang Dr.,

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
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  • Seamus J. Holden,

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
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  • Achillefs N. Kapanidis Dr.

    1. Biological Physics Research Group, Department of Physics, University of Oxford, Clarendon Laboratory, Parks Road, Oxford, OX1 3PU (United Kingdom)
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  • We thank Dr. M. Brenowitz (Albert Einstein College of Medicine) and Dr. R. Ebright (HHMI/Rutgers University) for plasmids, Dr. S. Weiss (UCLA) for software, and L. Sattary and J. Ghadiali for assistance. Funding was provided by the UK Bionanotechnology IRC, the EU (MIRG-CT-2005-031079), EPSRC (EP/D058775), and the Wellcome Trust (VS/06/OX/A4). M.H. was supported by a DAAD fellowship.

Abstract

original image

Transcription factors (TFs) control gene expression and are promising biomarkers for diseases. In a new, robust, and sensitive single-molecule fluorescence assay TFs and associated small molecules (such as nucleotides and sugars) are detected based on DNA coincidence of the two halves of the binding site (see picture; S=stoichiometry). The assay is compatible with measurements in solution, on solid supports, and even in complex biological samples.

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