This study investigates the role of G-quadruplex (G4) DNA structures in gene transcription. Researchers used a synthetic biology approach to integrate G4-forming sequences into the promoter of a synthetic reporter gene in human cells. They demonstrated that the formation of G4 structures in gene promoters enhances transcription, independent of primary sequence or inherent G-richness. Systematic variation in G4 stability showed that higher stability leads to increased transcriptional levels. The study also showed that G4 structures can stimulate transcription even when the SP1 consensus motif is absent, indicating that the folded G4 structure itself is the key factor in transcriptional activation. Additionally, the stability of G4 structures was found to influence their formation in cells, with more stable G4s leading to higher levels of transcription. These findings support the idea that G4 structures can act as positive regulators of transcription by recruiting proteins such as transcription factors and chromatin remodelers. The study highlights the importance of G4 structures in gene regulation and provides insights into their potential role in transcriptional control.This study investigates the role of G-quadruplex (G4) DNA structures in gene transcription. Researchers used a synthetic biology approach to integrate G4-forming sequences into the promoter of a synthetic reporter gene in human cells. They demonstrated that the formation of G4 structures in gene promoters enhances transcription, independent of primary sequence or inherent G-richness. Systematic variation in G4 stability showed that higher stability leads to increased transcriptional levels. The study also showed that G4 structures can stimulate transcription even when the SP1 consensus motif is absent, indicating that the folded G4 structure itself is the key factor in transcriptional activation. Additionally, the stability of G4 structures was found to influence their formation in cells, with more stable G4s leading to higher levels of transcription. These findings support the idea that G4 structures can act as positive regulators of transcription by recruiting proteins such as transcription factors and chromatin remodelers. The study highlights the importance of G4 structures in gene regulation and provides insights into their potential role in transcriptional control.