Single-molecule DNA-flow stretching assay as a versatile hybrid tool for investigating DNA-protein interactions
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Single-molecule techniques allow researchers to investigateindividual molecules and obtain unprecedented details of theheterogeneous nature of biological entities. They play instrumentalroles in studying DNA-protein interactions due to theability to visualize DNA or proteins and to manipulate individualDNA molecules by applying force or torque. Here, wedescribe single-molecule DNA-flow stretching assays as hybridtools that combine forces with fluorescence. We also reviewhow widely these assays are utilized in elucidating workingmechanisms of DNA-binding proteins. Additionally, we providea brief explanation of various efforts to prepare DNA substrateswith desired internal protein-binding sequences. More complicatedneeds for DNA-protein interaction research have led to improvementsin single-molecule DNA flow-stretching techniques. Several DNA flow-stretching variants such as DNA curtain,DNA motion capture assays, and protein-induced fluorescenceenhancement (PIFE) are introduced in this mini review. SinglemoleculeDNA flow-stretching assays will keep contributing toour understanding of how DNA-binding proteins function dueto their multiplexed, versatile, and robust capabilities.
Abstract
Single-molecule techniques allow researchers to investigateindividual molecules and obtain unprecedented details of theheterogeneous nature of biological entities. They play instrumentalroles in studying DNA-protein interactions due to theability to visualize DNA or proteins and to manipulate individualDNA molecules by applying force or torque. Here, wedescribe single-molecule DNA-flow stretching assays as hybridtools that combine forces with fluorescence. We also reviewhow widely these assays are utilized in elucidating workingmechanisms of DNA-binding proteins. Additionally, we providea brief explanation of various efforts to prepare DNA substrateswith desired internal protein-binding sequences. More complicatedneeds for DNA-protein interaction research have led to improvementsin single-molecule DNA flow-stretching techniques. Several DNA flow-stretching variants such as DNA curtain,DNA motion capture assays, and protein-induced fluorescenceenhancement (PIFE) are introduced in this mini review. SinglemoleculeDNA flow-stretching assays will keep contributing toour understanding of how DNA-binding proteins function dueto their multiplexed, versatile, and robust capabilities.