Illuminating nanoscale motion using single-molecule Förster resonance energy transfer: Latest insights and innovations

Bio-macromolecules rely on dynamic conformational changes for their function. While static 3D structures have provided invaluable insights into the spatial architecture of biomolecules, they lack temporal resolution. Complementarily, single molecule Förster resonance energy transfer (smFRET) enables real-time recording of nanoscale motions, providing a unique link between structure and function. This review highlights recent biological insights and methodological advances of the past two years. First, with examples from across the central dogma of molecular biology, we highlight mechanisms that became accessible through smFRET, complementing traditional structural biology techniques. Second, we review technological innovations, namely smFRET-specific progress, in situ integration of smFRET with other experiments, and ex situ complementary studies. Altogether, these advances demonstrate that the era of dynamic structural biology has truly arrived.