Single-Molecule Fluorescence Resonance Energy Transfer and Its Biomedical Applications

Abstract Single-molecule fluorescence resonance energy transfer (smFRET) requires significant spectral overlap between the donor emission and the acceptor absorption, and its efficiency is strongly dependent on the donor-acceptor distance. Consequently, smFRET has the capability to study molecular interactions of biomolecules with sub-nanometer resolution in a homogeneous manner without the involvement of any separation steps. The smFRET has significant advantages of high signal-to-noise ratio, low sample consumption, and homogeneous assay, providing a new approach to detect low-abundant biomolecules with extremely high sensitivity. In this review, we summarize the strategies for improving smFRET efficiency and the applications of smFRET in sensitive detection of DNAs, microRNAs, enzymes, and in situ imaging of single-nucleotide mutation. Moreover, we provide new insight into the challenges and future perspectives of smFRET.