Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers

Sischka A, Spiering A, Khaksar M, Laxa M, König J, Dietz K-J, Anselmetti D (2010)
JOURNAL OF PHYSICS-CONDENSED MATTER 22(45).

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Abstract
We investigated the threading and controlled translocation of individual lambda-DNA (lambda-DNA) molecules through solid-state nanopores with piconewton force sensitivity, millisecond time resolution and picoampere ionic current sensitivity with a set-up combining quantitative 3D optical tweezers (OT) with electrophysiology. With our virtually interference-free OT set-up the binding of RecA and single peroxiredoxin protein molecules to lambda-DNA was quantitatively investigated during dynamic translocation experiments where effective forces and respective ionic currents of the threaded DNA molecule through the nanopore were measured during inward and outward sliding. Membrane voltage-dependent experiments of reversible single protein/DNA translocation scans yield hysteresis-free, asymmetric single-molecule fingerprints in the measured force and conductance signals that can be attributed to the interplay of optical trap and electrostatic nanopore potentials. These experiments allow an exact localization of the bound protein along the DNA strand and open fascinating applications for label-free detection of DNA-binding ligands, where structural and positional binding phenomena can be investigated at a single-molecule level.
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Sischka A, Spiering A, Khaksar M, et al. Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers. JOURNAL OF PHYSICS-CONDENSED MATTER. 2010;22(45).
Sischka, A., Spiering, A., Khaksar, M., Laxa, M., König, J., Dietz, K. - J., & Anselmetti, D. (2010). Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers. JOURNAL OF PHYSICS-CONDENSED MATTER, 22(45).
Sischka, A., Spiering, A., Khaksar, M., Laxa, M., König, J., Dietz, K. - J., and Anselmetti, D. (2010). Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers. JOURNAL OF PHYSICS-CONDENSED MATTER 22.
Sischka, A., et al., 2010. Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers. JOURNAL OF PHYSICS-CONDENSED MATTER, 22(45).
A. Sischka, et al., “Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers”, JOURNAL OF PHYSICS-CONDENSED MATTER, vol. 22, 2010.
Sischka, A., Spiering, A., Khaksar, M., Laxa, M., König, J., Dietz, K.-J., Anselmetti, D.: Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers. JOURNAL OF PHYSICS-CONDENSED MATTER. 22, (2010).
Sischka, Andy, Spiering, André, Khaksar, Maryam, Laxa, Miriam, König, Janine, Dietz, Karl-Josef, and Anselmetti, Dario. “Dynamic translocation of ligand-complexed DNA through solid-state nanopores with optical tweezers”. JOURNAL OF PHYSICS-CONDENSED MATTER 22.45 (2010).
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5 Citations in Europe PMC

Data provided by Europe PubMed Central.

Nanopore sensing of individual transcription factors bound to DNA.
Squires A, Atas E, Meller A., Sci Rep 5(), 2015
PMID: 26109509
Electrically facilitated translocation of protein through solid nanopore.
Wu L, Liu H, Zhao W, Wang L, Hou C, Liu Q, Lu Z., Nanoscale Res Lett 9(1), 2014
PMID: 24661490
Video-based and interference-free axial force detection and analysis for optical tweezers.
Knust S, Spiering A, Vieker H, Beyer A, Golzhauser A, Tonsing K, Sischka A, Anselmetti D., Rev Sci Instrum 83(10), 2012
PMID: 23126771
Note: Direct force and ionic-current measurements on DNA in a nanocapillary.
Otto O, Steinbock LJ, Wong DW, Gornall JL, Keyser UF., Rev Sci Instrum 82(8), 2011
PMID: 21895280
Controlling molecular transport through nanopores.
Keyser UF., J R Soc Interface 8(63), 2011
PMID: 21715402

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