The study investigates the role of G-quadruplexes (G4s) in gene transcription. G4s are DNA structures that can form in the human genome, particularly in gene promoters, and are associated with active chromatin and transcription factor recruitment. The researchers used a synthetic biology approach to integrate G4-forming sequences into the promoter of a reporter gene in human cells. They found that these G4 structures within the promoter stimulate 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 demonstrated that the formation of G4 structures in a defined gene promoter can increase gene transcription, aligning with previous observations linking G4s to active transcription. The findings support the idea that G4 structures positively regulate gene expression by recruiting transcription factors and chromatin remodelers.The study investigates the role of G-quadruplexes (G4s) in gene transcription. G4s are DNA structures that can form in the human genome, particularly in gene promoters, and are associated with active chromatin and transcription factor recruitment. The researchers used a synthetic biology approach to integrate G4-forming sequences into the promoter of a reporter gene in human cells. They found that these G4 structures within the promoter stimulate 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 demonstrated that the formation of G4 structures in a defined gene promoter can increase gene transcription, aligning with previous observations linking G4s to active transcription. The findings support the idea that G4 structures positively regulate gene expression by recruiting transcription factors and chromatin remodelers.