Laboratoire D'Optique et Biosciences, Ecole Polytechnique, CNRS, Inserm, Institut Polytechnique de Paris, 91120, Palaiseau, France; CNRS UMR9187, INSERM U1196, Université Paris-Saclay, F-91405, Orsay, France.
CNRS UMR9187, INSERM U1196, Université Paris-Saclay, F-91405, Orsay, France; CNRS UMR9187, INSERM U1196, Institut Curie, PSL Research University, F-91405, Orsay, France.
Biochimie. 2023 Nov;214(Pt A):5-23. doi: 10.1016/j.biochi.2022.12.019. Epub 2022 Dec 31.
Besides the well-known DNA double-helix, non-canonical nucleic acid structures regulate crucial biological activities. Among these oddities, guanine-rich DNA sequences can form unusual four-stranded secondary structures called G-quadruplexes (G4s). G4-prone sequences have been found in the genomes of most species, and G4s play important roles in essential processes such as transcription, replication, genome integrity and epigenetic regulation. Here, we present a short overview of G-quadruplexes followed by a detailed description of the biophysical and biochemical methods used to characterize G4s in vitro. The principles, experimental details and possible shortcomings of each method are discussed to provide a comprehensive view of the techniques used to study these structures. We aim to provide a set of guidelines for standardizing research on G-quadruplexes; these guidelines are not meant to be a dogmatic set of rules, but should rather provide useful information on the methods currently used to study these fascinating motifs.
除了众所周知的 DNA 双螺旋结构外，非典型核酸结构还调节着关键的生物活性。在这些奇异结构中，富含鸟嘌呤的 DNA 序列可以形成一种称为 G-四链体（G4s）的特殊四链二级结构。富含 G4 的序列已在大多数物种的基因组中被发现，G4 在转录、复制、基因组完整性和表观遗传调控等重要过程中发挥着重要作用。本文首先概述了 G-四链体，然后详细描述了用于体外鉴定 G4 的生物物理和生化方法。讨论了每种方法的原理、实验细节和可能的缺点，为研究这些结构的技术提供了全面的视角。我们旨在为 G-四链体的研究提供一套标准化指南；这些指南并非一套死板的规则，而是应该为目前用于研究这些迷人基序的方法提供有用的信息。
