Förster resonance energy transfer from Hoechst 33258 to SYBR Green I in cholesteric liquid-crystalline DNA

Abstract

In the present work Förster resonance energy transfer (FRET) between DNA-specific fluorescent dyes Hoechst 33258 (Ht58; donor) and SYBR Green I (SG; acceptor) is compared in isotropic DNA solution and in its cholesteric liquid-crystalline dispersion (DNA CLCD). At a fixed donor concentration of 3.57 × 10−6 М in the spatially ordered system, FRET starts at lower acceptor concentration than in the isotropic system and at the SG concentration below 1.05 × 10−7 M, its efficiency in DNA CLCD is up to ≈4-fold higher than in DNA solution (3D-FRET vs. 1D-FRET). However, the maximum FRET efficiency is higher in isotropic solution (≈70% vs. ≈50%, respectively) apparently due to the absence of steric restrictions on intermolecular interactions imposed by the dense packaging of the DNA molecules. In isotropic DNA solution FRET from Ht58 to SG is accompanied by the displacement of Ht58 from the minor-groove complex as the SG concentration increased, while the displacement of Ht58 in DNA CLCD seems to be limited. Using FRET between Ht58 and SG, the formation of DNA CLCD particles via stopped-flow method was investigated and for the first time the subsecond stage of the DNA CLCD particle formation, generation of “precholesteric” optically inactive nuclei, was demonstrated.