Journal Article

Locked nucleic acid oligomers as handles for single molecule manipulation

John P. Berezney and Omar A. Saleh

in Nucleic Acids Research

Volume 42, issue 19, pages e150-e150
Published in print October 2014 | ISSN: 0305-1048
Published online August 2014 | e-ISSN: 1362-4962 | DOI: http://dx.doi.org/10.1093/nar/gku760

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Single-molecule manipulation (SMM) techniques use applied force, and measured elastic response, to reveal microscopic physical parameters of individual biomolecules and details of biomolecular interactions. A major hurdle in the application of these techniques is the labeling method needed to immobilize biomolecules on solid supports. A simple, minimally-perturbative labeling strategy would significantly broaden the possible applications of SMM experiments, perhaps even allowing the study of native biomolecular structures. To accomplish this, we investigate the use of functionalized locked nucleic acid (LNA) oligomers as biomolecular handles that permit sequence-specific binding and immobilization of DNA. We find these probes form bonds with DNA with high specificity but with varied stability in response to the direction of applied mechanical force: when loaded in a shear orientation, the bound LNA oligomers were measured to be two orders of magnitude more stable than when loaded in a peeling, or unzipping, orientation. Our results show that LNA provides a simple, stable means to functionalize dsDNA for manipulation. We provide design rules that will facilitate their use in future experiments.

Journal Article.  4321 words.  Illustrated.

Subjects: Research Methods in Life Sciences

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