Human telomeric hybrid-2-over-hybrid-1 G-quadruplex targeting and a selective hypersaline-tolerant sensor using abasic site-engineered monomorphism

Abstract

Coexistence of the polymorphic hybrid-1 and hybrid-2 conformers for a given human telomeric G-quadruplex-forming sequence (htG4) complicates its fine structure identification and limits its application as a sensor element. With help from abasic site (AP site)-engineered htG4s serving as the monomorphic representatives of the two typical hybrid conformers, we found that thioflavin T (ThT) can selectively target the hybrid-2 conformer over the hybrid-1 counterpart in monomer and tandem htG4 molecules. The htG4 that solely adopts the monomorphic hybrid-2 conformer engineered by the AP site is most efficient in lighting up ThT fluorescence in K+ and a selective K+ sensor is realized with a remarkable hypersaline-tolerant capability that can work even in 30000-fold excess of Na+. At 600 mM Na+, the dynamic range for K+ detection can be extended to 30 mM with the limit of detection of 20 μM. This is the first report on the fluorescence discrimination of these two hybrid conformers of htG4 although they have long been categorized with their characteristically structural topologies. Our work will attract much interest in the development of sensors based on the monomorphic htG4 conformer since such high performance in sensor development has not been previously achieved.