研究人员介绍了一种高通量单分子并行分析方法，用于快速探索序列空间（SPARXS），将单分子荧光技术与下一代测序技术相结合。研究人员应用SPARXS揭示了Holliday接头的序列依赖性动力学，这是一种在同源重组中至关重要的中间体。
通过研究数百万个Holliday接头的动态，涵盖了数千种不同的序列，研究人员展示了SPARXS揭示序列模式、评估序列基序和构建热力学模型的能力。SPARXS作为一种多功能工具，能够解开分子尺度上序列特异性过程的机制。
据悉，分子生物学的核心在于序列、结构和功能之间的复杂相互作用。单分子技术提供了对结构和功能的深入动态洞察，但繁琐的检测方法限制了对大型序列库的功能筛选。
附：英文原文
Title: Single-molecule structural and kinetic studies across sequence space
Author: Ivo Severins, Carolien Bastiaanssen, Sung Hyun Kim, Roy B. Simons, John van Noort, Chirlmin Joo
Issue&Volume: 2024-08-23
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.
DOI: adn5968
Source: https://www.science.org/doi/10.1126/science.adn5968