High Resolution Optical Tweezers for Single-Molecule Studies of Eukaryotic Transcription
Investigating mechanical aspects of single RNA polymerases will further our understanding of the molecular mechanism of transcription elongation.
For these single-molecule experiments we establish a high resolution optical tweezers setup in the dual trap design, where one trap can be moved by a piezo-driven mirror. Experiments with 5 kbp DNA tethers attached to trapped beads have shown that our current resolution is about 0.3 nm for forces measured around 15 pN at a time resolution of approximately 0.1s. This is sufficient to study single base pair steps of RNA Polymerase II on DNA.
In further experiments we will investigate the behavior of RNA Polymerases I and II from a mechanical perspective. In addition, we will study transcription regulation by performing experiments with RNA Polymerase II in presence of different transcription factors such as TFIIS and TFIIF.
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This poster was presented at the 10th International Symposium on Scanning Probe Microscopy & Optical Tweezers in Life Science, 6-7 October 2011
K. Paul, A. Muschielok, N. M. Porcellato and J. Michaelis
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