Single-molecule structural and kinetic studies across sequence space
Ivo Severins (Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab, Universiteit Leiden)
Carolien Bastiaanssen (Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab)
Sung Hyun Kim (Ewha Womans University, Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab)
Roy B. Simons (Erasmus MC)
John Van Noort (Universiteit Leiden)
C Joo (Kavli institute of nanoscience Delft, TU Delft - BN/Chirlmin Joo Lab, Ewha Womans University)
More Info
expand_more
Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons.
Abstract
At the core of molecular biology lies the intricate interplay between sequence, structure, and function. Single-molecule techniques provide in-depth dynamic insights into structure and function, but laborious assays impede functional screening of large sequence libraries. We introduce high-throughput Single-molecule Parallel Analysis for Rapid eXploration of Sequence space (SPARXS), integrating single-molecule fluorescence with next-generation sequencing. We applied SPARXS to study the sequence-dependent kinetics of the Holliday junction, a critical intermediate in homologous recombination. By examining the dynamics of millions of Holliday junctions, covering thousands of distinct sequences, we demonstrated the ability of SPARXS to uncover sequence patterns, evaluate sequence motifs, and construct thermodynamic models. SPARXS emerges as a versatile tool for untangling the mechanisms that underlie sequence-specific processes at the molecular scale.