Correlating fluorescence microscopy, optical and magnetic tweezers to study single chiral biopolymers such as DNA

关联荧光显微镜、光镊和磁镊来研究 DNA 等单一手性生物聚合物

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作者:Jack W Shepherd #, Sebastien Guilbaud #, Zhaokun Zhou #, Jamieson A L Howard #, Matthew Burman, Charley Schaefer, Adam Kerrigan, Clare Steele-King, Agnes Noy, Mark C Leake

Abstract

Biopolymer topology is critical for determining interactions inside cell environments, exemplified by DNA where its response to mechanical perturbation is as important as biochemical properties to its cellular roles. The dynamic structures of chiral biopolymers exhibit complex dependence with extension and torsion, however the physical mechanisms underpinning the emergence of structural motifs upon physiological twisting and stretching are poorly understood due to technological limitations in correlating force, torque and spatial localization information. We present COMBI-Tweez (Combined Optical and Magnetic BIomolecule TWEEZers), a transformative tool that overcomes these challenges by integrating optical trapping, time-resolved electromagnetic tweezers, and fluorescence microscopy, demonstrated on single DNA molecules, that can controllably form and visualise higher order structural motifs including plectonemes. This technology combined with cutting-edge MD simulations provides quantitative insight into complex dynamic structures relevant to DNA cellular processes and can be adapted to study a range of filamentous biopolymers.

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