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Measuring Single-Molecule Twist and Torque in Multiplexed Magnetic Tweezers

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Book cover Nanoscale Imaging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1814))

Abstract

Magnetic tweezers permit application of precisely calibrated stretching forces to nucleic acid molecules tethered between a surface and superparamagnetic beads. In addition, magnetic tweezers can control the tethers’ twist. Here, we focus on recent extensions of the technique that expand the capabilities of conventional magnetic tweezers by enabling direct measurements of single-molecule torque and twist. Magnetic torque tweezers (MTT) still control the DNA or RNA tether’s twist, but directly measure molecular torque by monitoring changes in the equilibrium rotation angle upon overwinding and underwinding of the tether. In freely orbiting magnetic tweezers (FOMT), one end of the tether is allowed to rotate freely, while still applying stretching forces and monitoring rotation angle. Both MTT and FOMT have provided unique insights into the mechanical properties, structural transitions, and interactions of DNA and RNA. Here, we provide step-by-step protocols to carry out FOMT and MTT measurements. In particular, we focus on multiplexed measurements, i.e., measurements that record data for multiple nucleic acid tethers at the same time, to improve statistics and to facilitate the observation of rare events.

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Acknowledgments

We thank Jelle van der Does for help with instrument development, Susanne Hage for help with development of the DNA construct, Philipp Walker for instrument construction and useful discussions, the Rief chair at the TU Munich for use of the laser cutter, and the German Research Foundation (DFG) via Sonderforschungbereich SFB 863 “Forces in Biomolecular Systems” for funding.

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Authors and Affiliations

  1. Department of Physics, Nanosystems Initiative Munich, and Center for Nanoscience, LMU Munich, Munich, Germany

    Franziska Kriegel, Willem Vanderlinden, Thomas Nicolaus, Angelika Kardinal & Jan Lipfert

  2. Division of Molecular Imaging and Photonics, Department of Chemistry, KU Leuven—University of Leuven, Leuven, Belgium

    Willem Vanderlinden

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  1. Franziska Kriegel
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  2. Willem Vanderlinden
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  3. Thomas Nicolaus
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  4. Angelika Kardinal
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  5. Jan Lipfert
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Corresponding author

Correspondence to Jan Lipfert .

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Editors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Nebraska Medical Center, Omaha, Nebraska, USA

    Yuri L. Lyubchenko

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Kriegel, F., Vanderlinden, W., Nicolaus, T., Kardinal, A., Lipfert, J. (2018). Measuring Single-Molecule Twist and Torque in Multiplexed Magnetic Tweezers. In: Lyubchenko, Y. (eds) Nanoscale Imaging. Methods in Molecular Biology, vol 1814. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8591-3_6

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  • DOI: https://doi.org/10.1007/978-1-4939-8591-3_6

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