biostudies-literatureUnknown10(3)Parada ZGuanine-rich sequences exhibit a high degree of polymorphism and can form single-stranded, Watson-Crick duplex, and four-stranded G-quadruplex structures. These sequences have found a wide range of uses in synthetic biology applications, arising in part from their structural plasticity. High-throughput, low-cost tools for monitoring the folding and unfolding transitions of G-rich sequences would provide an enabling technology for accelerating the prototyping of synthetic biological systems and for accelerating design-build-test cycles. Here, we show that unfolding transitions of a range of G-quadruplex-forming DNA sequences can be monitored in a FRET-like format using DNA sequences that possess only a single dye label, with no quencher. These quencher-free assays can be performed at low coACS omega3176-3181https://www.ebi.ac.uk/biostudies/studies/S-EPMC11780409biostudies-literatureQuencher-Free Fluorescence Monitoring of G-Quadruplex Folding.PMC11780409Adamala KPHoog TGParada ZEngelhart AEfalseQuencher-Free Fluorescence Monitoring of G-Quadruplex Folding.Guanine-rich sequences exhibit a high degree of polymorphism and can form single-stranded, Watson-Crick duplex, and four-stranded G-quadruplex structures. These sequences have found a wide range of uses in synthetic biology applications, arising in part from their structural plasticity. High-throughput, low-cost tools for monitoring the folding and unfolding transitions of G-rich sequences would provide an enabling technology for accelerating the prototyping of synthetic biological systems and for accelerating design-build-test cycles. Here, we show that unfolding transitions of a range of G-quadruplex-forming DNA sequences can be monitored in a FRET-like format using DNA sequences that possess only a single dye label, with no quencher. These quencher-free assays can be performed at low co2025-01-01T00:00:00Z2025 Jan2025-04-03T23:26:39.119Z2025-04-03T23:26:39.119ZS-EPMC117804093989573310.1021/acsomega.4c10720