Alanazi, Abeer FR;
Haider, Shozeb;
Parkinson, Gary N;
(2024)
The use of single-molecule FRET for the characterization of Holliday junctions containing human telomeric DNA: a methodological approach for nanoscale distance, mobility, and stability measurements.
Medicinal Chemistry Research
, 33
pp. 2066-2078.
10.1007/s00044-024-03197-0.
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Abstract
Single-molecule analysis has emerged as a powerful biophysical technique, allowing the investigation of molecular interactions at unprecedented spatial resolutions. In this paper, we outline a step-by-step procedure for the design of DNA sequences to form Holliday junction structures containing repetitive telomeric sequences and exploring conformational dynamics using Single-Molecule Förster Resonance Energy Transfer (smFRET). We describe in detail the setup and optimization of the single-molecule fluorescence microscope, along with data acquisition and analysis procedures. Furthermore, we address potential challenges and provide troubleshooting strategies to ensure successful data collection and interpretation. The presented methodology serves as a valuable resource for researchers aiming to employ smFRET to study Holliday junctions and other biomolecular systems at the nanoscale, paving the way for further advancements in the field of single-molecule biophysics. (Figure presented.)
| Type: | Article |
|---|---|
| Title: | The use of single-molecule FRET for the characterization of Holliday junctions containing human telomeric DNA: a methodological approach for nanoscale distance, mobility, and stability measurements |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1007/s00044-024-03197-0 |
| Publisher version: | http://dx.doi.org/10.1007/s00044-024-03197-0 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Nanoimaging, DNA, Chromophores, Single molecules |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy > Pharma and Bio Chemistry |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10189801 |
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