Biophysical Society Thematic Meeting | Tutzing 2026

Single-Molecule FRET: The Next 30 Years

Friday Speaker Abstracts

FROM SEQUENCE TO FUNCTION: BRIDGING SINGLE-MOLECULE KINETICS WITH MOLECULAR IDENTITY Sebastian Deindl 1 ; 1 Uppsala Biomedical Center, University of Uppsala, Uppsala, Sweden Fundamental biological processes rely on macromolecules that often exhibit complex dynamics tightly linked to their functions. Single-molecule techniques are ideally suited to characterize these dynamics but are typically limited to probing only a small number of different samples. However, a comprehensive understanding of complex biological systems frequently requires the analysis of a large number of samples. For example, studying nucleic acid-interacting enzymes often demands exploration across a broad sequence space. I will introduce our recently developed method, MUSCLE (MUltiplexed Single-molecule Characterization at the Library scalE), which combines single-molecule fluorescence microscopy with next-generation sequencing. This approach enables highly multiplexed observations of the dynamics of millions of individual molecules across thousands of distinct samples. Using MUSCLE, we comprehensively profiled the sequence dependence of DNA hairpin properties and the dynamics of Cas9-induced target DNA unwinding and rewinding. I will also present our recent applications of MUSCLE to investigate how sequence context influences the dynamics of DNA mismatch repair and its functional outcomes.

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