Force-fluorescence setup for observing protein-DNA interactions under load

Abstract

This chapter explores advanced single-molecule techniques for studying protein-DNA interactions, particularly focusing on Replication Protein A (RPA) using a force-fluorescence setup. It combines magnetic tweezers (MT) with total internal reflection fluorescence (TIRF) microscopy, enabling detailed observation of DNA behavior under mechanical stress. The chapter details the use of DNA hairpins and bare DNA to examine RPA's binding dynamics and its influence on DNA's mechanical properties. This approach provides deeper insights into RPA's role in DNA replication, repair, and recombination, highlighting its significance in maintaining genomic stability.

Keywords: DNA hairpins; DNA overstretching; Magnetic tweezers (MT); Protein–DNA interaction; Replication protein A (RPA); Total internal reflection fluorescence (TIRF) microscopy.