Microtech 2011

Lighting up Individual DNA Binding Proteins with Quantum Dots

Y. Ebenstein, S. Kim, R. Lin, Y. Kim, N. Gassman, X. Michalet, S. Weiss

Keywords: optical mapping, quantum dots, Epigenetics


The ability to determine the precise loci and occupancy of DNA-binding proteins is instrumental to our understanding of cellular processes like gene expression and regulation. We propose a single-molecule approach for the direct visualization of proteins bound to their template DNA. Fluorescent quantum-dots (QD) are used to label proteins bound to DNA, allowing multicolor, nm-resolution localization. Protein-DNA complexes are linearly extended and imaged to determine the precise location of the protein binding sites. The method is demonstrated by detecting individual QD-labeled T7-RNA polymerases on the T7 bacteriophage genome. This work demonstrates the potential of this approach to precisely read DNA binding protein position information essential for both basic science and medical diagnostics applications.

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