Function through shape: An overview of DNA G-quadruplexes in transcriptional regulation

G-quadruplexes (G4s) are four-stranded nucleic acid structures formed by guanine-rich sequences. Over the past two decades, G4s have emerged as key elements in genome organisation and transcription. Their remarkable structural diversity distinguishes G4s from other non-canonical DNA structures, including i-motifs and triplexes. Recent evidence suggests that G4s may serve as emerging regulatory hubs for transcriptional control, adding complexity to simple models in which G4s act solely as recruiters of transcription factors at gene promoters. Emerging data indicate that G4s can influence nucleosome occupancy, chromatin accessibility, and long-range chromatin interactions. Recent evidence also suggests a key role of G4s in modulating the formation of biomolecular condensate, which have been widely implicated in the regulation of transcription and chromatin organisation. Altogether, these findings highlight the potential of G4s to act as architectural elements of chromatin. In this review, we examine what makes G4s structurally unique compared to other alternative DNA structures, and discuss their potential involvement in genome architecture, nucleosome occupancy, and chromatin looping. We also highlight recent methodological advances, from small molecule stabilisers to CRISPR-based precision targeting, that enabled the manipulation of individual G4s in their native genomic context, revealing context-dependent G4 functions and highlighting their potential as therapeutic targets.