Nastaran Hadizadeh (LUMICKS)| Jason Lin (LUMICKS)| Trey Simpson (LUMICKS)| Rosalie Driessen (LUMICKS)| Aida Llauró-Portell (LUMICKS)| Andrea Candelli (LUMICKS)
Biological processes performed by proteins interacting with DNA and chromatin are key to cell health and viability. Detailed insights into these processes provide essential information for understanding the pathological conditions that develop when such processes go awry and provide us with tools to alter the fate of the cells. Direct, real-time observations of individual proteins interacting with DNA are required to validate and complete the current biological models. Single-molecule technologies offer an exciting opportunity to meet these challenges and to study protein function and activity in real-time. Here, we present our efforts for further enabling discoveries in the field of DNA-protein interactions using both the combination of optical tweezers with correlative confocal fluorescence microscopy. We present several examples in which our technologies enhanced the understanding of the DNA repair mechanisms, chromatin structure and DNA editing tools. Furthermore, we show that advances in hybrid single-molecule methods can be turned into an easy-to-use and stable instrument that has the ability to open up new venues in the field of DNA-protein interactions.
Keywords: single-molecule, DNA-protein interaction, chromatin, DNA editing, CRISPR Cas9, DNA repair, Rad 51